Characterizing the insecticide resistance of Anopheles gambiae in Mali.

PubMed

Cisse, Moussa B M; Keita, Chitan; Dicko, Abdourhamane; Dengela, Dereje; Coleman, Jane; Lucas, Bradford; Mihigo, Jules; Sadou, Aboubacar; Belemvire, Allison; George, Kristen; Fornadel, Christen; Beach, Raymond

2015-08-22

The impact of indoor residual spraying (IRS) and long-lasting insecticide nets (LLINs), key components of the national malaria control strategy of Mali, is threatened by vector insecticide resistance. The objective of this study was to assess the level of insecticide resistance in Anopheles gambiae sensu lato populations from Mali against four classes of insecticide recommended for IRS: organochlorines (OCs), pyrethroids (PYs), carbamates (CAs) and organophosphates (OPs). Characterization of resistance was done in 13 sites across southern Mali and assessed presence and distribution of physiological mechanisms that included target-site modifications: knockdown resistance (kdr) and altered acetycholinesterase (AChE), and/or metabolic mechanisms: elevated esterases, glutathione S-transferases (GSTs), and monooxygenases. The World Health Organization (WHO) tube test was used to determine phenotypic resistance of An. gambiae s.l. to: dichlorodiphenyltrichloroethane (DDT) (OC), deltamethrin (PY), lambda-cyhalothrin (PY), bendiocarb (CA), and fenitrothion (OP). Identification of sibling species and presence of the ace-1 (R) and Leu-Phe kdr, resistance-associated mutations, were determined using polymerase chain reaction (PCR) technology. Biochemical assays were conducted to detect increased activity of GSTs, oxidases and esterases. Populations tested showed high levels of resistance to DDT in all 13 sites, as well as increased resistance to deltamethrin and lambda-cyhalothrin in 12 out of 13 sites. Resistance to fenitrothion and bendiocarb was detected in 1 and 4 out of 13 sites, respectively. Anopheles coluzzii, An. gambiae sensu stricto and Anopheles arabiensis were identified with high allelic frequencies of kdr in all sites where each of the species were found (13, 12 and 10 sites, respectively). Relatively low allelic frequencies of ace-1 (R) were detected in four sites where this assessment was conducted. Evidence of elevated insecticide metabolism, based on oxidase

Insecticide susceptibility status of Anopheles gambiae s.l. (Diptera: Culicidae) in the Republic of Cameroon.

PubMed

Etang, Josiane; Manga, Lucien; Chandre, Fabrice; Guillet, Pierre; Fondjo, Etienne; Mimpfoundi, Remy; Toto, Jean-Claude; Fontenille, Didier

2003-07-01

A large-scale survey of Anopheles gambiae Giles, 1902 susceptibility to DDT, dieldrin, permethrin, and deltamethrin was conducted in the Republic of Cameroon. 15 field populations from various geographical areas were tested using World Health Organization test kits for adult mosquitoes. The laboratory Kisumu susceptible reference strain was tested as a control. Results showed that dieldrin and DDT resistance was still present in some populations, and indicated permethrin or deltamethrin resistance. Within the Anopheles gambiae complex, resistant individuals belonged to An. gambiae s.s. and An. arabiensis species. Both M and S molecular forms of An. gambiae s.s. were found resistant. In most of resistant populations, the knockdown times were 2-5-folds increased. However, none of the surviving mosquitoes was positive to the kdr "Leu-Phe" mutation using polymerase chain reaction (PCR) diagnostic test. These results likely suggested involvement of other resistance mechanism(s), such as enzyme detoxification or kdr "Leu-Ser" mutation. Researches on An. gambiae s.l. resistance should be promoted in Cameroon, to improve malaria vector control programs and to implement resistance management strategies.

G Protein-Coupled Receptors in Anopheles gambiae

NASA Astrophysics Data System (ADS)

Hill, Catherine A.; Fox, A. Nicole; Pitts, R. Jason; Kent, Lauren B.; Tan, Perciliz L.; Chrystal, Mathew A.; Cravchik, Anibal; Collins, Frank H.; Robertson, Hugh M.; Zwiebel, Laurence J.

2002-10-01

We used bioinformatic approaches to identify a total of 276 G protein-coupled receptors (GPCRs) from the Anopheles gambiae genome. These include GPCRs that are likely to play roles in pathways affecting almost every aspect of the mosquito's life cycle. Seventy-nine candidate odorant receptors were characterized for tissue expression and, along with 76 putative gustatory receptors, for their molecular evolution relative to Drosophila melanogaster. Examples of lineage-specific gene expansions were observed as well as a single instance of unusually high sequence conservation.

Behavioural response of the malaria vector Anopheles gambiae to host plant volatiles and synthetic blends

USDA-ARS?s Scientific Manuscript database

Sugar feeding is critical for survival of malaria vectors and, although discriminative plant feeding previously has been shown to occur in Anopheles gambiae s.s., little is known about the cues mediating attraction to these plants. In this study, we investigated the role of olfaction in An. gambiae ...

Molasses as a source of carbon dioxide for attracting the malaria mosquitoes Anopheles gambiae and Anopheles funestus

PubMed Central

2014-01-01

Background Most odour baits for haematophagous arthropods contain carbon dioxide (CO2). The CO2 is sourced artificially from the fermentation of refined sugar (sucrose), dry ice, pressurized gas cylinders or propane. These sources of CO2 are neither cost-effective nor sustainable for use in remote areas of sub-Saharan Africa. In this study, molasses was evaluated as a potential substrate for producing CO2 used as bait for malaria mosquitoes. Methods The attraction of laboratory-reared and wild Anopheles gambiae complex mosquitoes to CO2 generated from yeast-fermentation of molasses was assessed under semi-field and field conditions in western Kenya. In the field, responses of wild Anopheles funestus were also assessed. Attraction of the mosquitoes to a synthetic mosquito attractant, Mbita blend (comprising ammonia, L-lactic acid, tetradecanoic acid and 3-methyl-1-butanol) when augmented with CO2 generated from yeast fermentation of either molasses or sucrose was also investigated. Results In semi-field, the release rate of CO2 and proportion of An. gambiae mosquitoes attracted increased in tandem with an increase in the quantity of yeast-fermented molasses up to an optimal ratio of molasses and dry yeast. More An. gambiae mosquitoes were attracted to a combination of the Mbita blend plus CO2 produced from fermenting molasses than the Mbita blend plus CO2 from yeast-fermented sucrose. In the field, significantly more female An. gambiae sensu lato mosquitoes were attracted to the Mbita blend augmented with CO2 produced by fermenting 500 g of molasses compared to 250 g of sucrose or 250 g of molasses. Similarly, significantly more An. funestus, Culex and other anopheline mosquito species were attracted to the Mbita blend augmented with CO2 produced from fermenting molasses than the Mbita blend with CO2 produced from sucrose. Augmenting the Mbita blend with CO2 produced from molasses was associated with high catches of blood-fed An. gambiae s.l. and An. funestus

Odor Coding in the Maxillary Palp of the Malaria Vector Mosquito Anopheles gambiae

PubMed Central

Lu, Tan; Qiu, Yu Tong; Wang, Guirong; Kwon, Jae Young; Rutzler, Michael; Kwon, Hyung-Wook; Pitts, R. Jason; van Loon, Joop J.A.; Takken, Willem; Carlson, John R.; Zwiebel, Laurence J.

2011-01-01

Summary Background Many species of mosquitoes, including the major malaria vector Anopheles gambiae, utilize carbon dioxide (CO2) and 1-octen-3-ol as olfactory cues in host-seeking behaviors that underlie their vectorial capacity. However, the molecular and cellular basis of such olfactory responses remains largely unknown. Results Here, we use molecular and physiological approaches coupled with systematic functional analyses to define the complete olfactory sensory map of the An. gambiae maxillary palp, an olfactory appendage that mediates the detection of these compounds. In doing so, we identify three olfactory receptor neurons (ORNs) that are organized in stereotyped triads within the maxillary-palp capitate-peg-sensillum population. One ORN is CO2-responsive and characterized by the coexpression of three receptors that confer CO2 responses, whereas the other ORNs express characteristic odorant receptors (AgORs) that are responsible for their in vivo olfactory responses. Conclusions Our results describe a complete and highly concordant map of both the molecular and cellular olfactory components on the maxillary palp of the adult female An. gambiae mosquito. These results also facilitate the understanding of how An. gambiae mosquitoes sense olfactory cues that might be exploited to compromise their ability to transmit malaria. PMID:17764944

Breakpoint structure of the Anopheles gambiae 2Rb chromosomal inversion.

PubMed

Lobo, Neil F; Sangaré, Djibril M; Regier, Allison A; Reidenbach, Kyanne R; Bretz, David A; Sharakhova, Maria V; Emrich, Scott J; Traore, Sekou F; Costantini, Carlo; Besansky, Nora J; Collins, Frank H

2010-10-25

Alternative arrangements of chromosome 2 inversions in Anopheles gambiae are important sources of population structure, and are associated with adaptation to environmental heterogeneity. The forces responsible for their origin and maintenance are incompletely understood. Molecular characterization of inversion breakpoints provides insight into how they arose, and provides the basis for development of molecular karyotyping methods useful in future studies. Sequence comparison of regions near the cytological breakpoints of 2Rb allowed the molecular delineation of breakpoint boundaries. Comparisons were made between the standard 2R+b arrangement in the An. gambiae PEST reference genome and the inverted 2Rb arrangements in the An. gambiae M and S genome assemblies. Sequence differences between alternative 2Rb arrangements were exploited in the design of a PCR diagnostic assay, which was evaluated against the known chromosomal banding pattern of laboratory colonies and field-collected samples from Mali and Cameroon. The breakpoints of the 7.55 Mb 2Rb inversion are flanked by extensive runs of the same short (72 bp) tandemly organized sequence, which was likely responsible for chromosomal breakage and rearrangement. Application of the molecular diagnostic assay suggested that 2Rb has a single common origin in An. gambiae and its sibling species, Anopheles arabiensis, and also that the standard arrangement (2R+b) may have arisen twice through breakpoint reuse. The molecular diagnostic was reliable when applied to laboratory colonies, but its accuracy was lower in natural populations. The complex repetitive sequence flanking the 2Rb breakpoint region may be prone to structural and sequence-level instability. The 2Rb molecular diagnostic has immediate application in studies based on laboratory colonies, but its usefulness in natural populations awaits development of complementary molecular tools.

Evaluating preservation methods for identifying Anopheles gambiae s.s. and Anopheles arabiensis complex mosquitoes species using near infra-red spectroscopy.

PubMed

Mayagaya, Valeriana Simon; Ntamatungiro, Alex John; Moore, Sarah Jane; Wirtz, Robert Andrew; Dowell, Floyd Ercell; Maia, Marta Ferreira

2015-01-27

Near-infrared spectroscopy (NIRS) has been successfully used on fresh and RNAlater-preserved members of the Anopheles gambiae complex to identify sibling species and age. No preservation methods other than using RNAlater have been tested to preserve mosquitoes for species identification using NIRS. However, RNAlater is not the most practical preservative for field settings because it is expensive, requires basic laboratory conditions for storage and is not widely available in sub-Saharan Africa. The aim of this study was to test several cheaper and more field-friendly preservation methods for identifying sibling species of the An. gambiae complex using NIRS. In this study we describe the use of NIRS to identify sibling species of preserved An. gambiae s. s. and An. arabiensis. Mosquitoes of each species were placed in sample tubes and preserved using one of the following preservation methods: (i) refrigeration at 4°C, (ii) freezing at -20°C, (iii) drying over a silica-gel desiccant, (iv) submersion in RNAlater at room temperature, (v) submersion in RNAlater at 4°C, and (vi) submersion in RNAlater at -20°C. Mosquitoes were preserved for 1, 4, 10, 32 or 50 weeks before they were scanned. Storage at 4°C was the only preservation method that, up to 32 weeks, did not result in significantly lower predicted values than those obtained from fresh insects. After 50 weeks, however, refrigerated samples did not give meaningful results. When storing for 50 weeks, desiccating samples over silica gel was the best preservation method, with a partial least squares regression cross-validation of >80%. Predictive data values were analyzed using a generalized linear model. NIRS can be used to identify species of desiccated Anopheles gambiae s.s. and Anopheles arabiensis for up to 50 weeks of storage with more than 80% accuracy.

The Anopheles gambiae transcriptome - a turning point for malaria control.

PubMed

Domingos, A; Pinheiro-Silva, R; Couto, J; do Rosário, V; de la Fuente, J

2017-04-01

Mosquitoes are important vectors of several pathogens and thereby contribute to the spread of diseases, with social, economic and public health impacts. Amongst the approximately 450 species of Anopheles, about 60 are recognized as vectors of human malaria, the most important parasitic disease. In Africa, Anopheles gambiae is the main malaria vector mosquito. Current malaria control strategies are largely focused on drugs and vector control measures such as insecticides and bed-nets. Improvement of current, and the development of new, mosquito-targeted malaria control methods rely on a better understanding of mosquito vector biology. An organism's transcriptome is a reflection of its physiological state and transcriptomic analyses of different conditions that are relevant to mosquito vector competence can therefore yield important information. Transcriptomic analyses have contributed significant information on processes such as blood-feeding parasite-vector interaction, insecticide resistance, and tissue- and stage-specific gene regulation, thereby facilitating the path towards the development of new malaria control methods. Here, we discuss the main applications of transcriptomic analyses in An. gambiae that have led to a better understanding of mosquito vector competence. © 2017 The Royal Entomological Society.

Development of a Gravid Trap for Collecting Live Malaria Vectors Anopheles gambiae s.l.

PubMed Central

Dugassa, Sisay; Lindh, Jenny M.; Oyieke, Florence; Mukabana, Wolfgang R.; Lindsay, Steven W.; Fillinger, Ulrike

2013-01-01

Background Effective malaria vector control targeting indoor host-seeking mosquitoes has resulted in fewer vectors entering houses in many areas of sub-Saharan Africa, with the proportion of vectors outdoors becoming more important in the transmission of this disease. This study aimed to develop a gravid trap for the outdoor collection of the malaria vector Anopheles gambiae s.l. based on evaluation and modification of commercially available gravid traps. Methods Experiments were implemented in an 80 m2 semi-field system where 200 gravid Anopheles gambiae s.s. were released nightly. The efficacy of the Box, CDC and Frommer updraft gravid traps was compared. The Box gravid trap was tested to determine if the presence of the trap over water and the trap’s sound affected catch size. Mosquitoes approaching the treatment were evaluated using electrocuting nets or detergents added to the water in the trap. Based on the results, a new gravid trap (OviART trap) that provided an open, unobstructed oviposition site was developed and evaluated. Results Box and CDC gravid traps collected similar numbers (relative rate (RR) 0.8, 95% confidence interval (CI) 0.6–1.2; p = 0.284), whereas the Frommer trap caught 70% fewer mosquitoes (RR 0.3, 95% CI 0.2–0.5; p < 0.001). The number of mosquitoes approaching the Box trap was significantly reduced when the trap was positioned over a water-filled basin compared to an open pond (RR 0.7 95% CI 0.6–0.7; p < 0.001). This effect was not due to the sound of the trap. Catch size increased by 60% (RR 1.6, 1.2–2.2; p = 0.001) with the new OviART trap. Conclusion Gravid An. Gambiae s.s. females were visually deterred by the presence of the trapping device directly over the oviposition medium. Based on these investigations, an effective gravid trap was developed that provides open landing space for egg-laying Anopheles . PMID:23861952

Sampling Outdoor, Resting Anopheles gambiae and Other Mosquitoes (Diptera: Culicidae) in Western Kenya with Clay Pots

PubMed Central

Odiere, M.; Bayoh, M. N.; Gimnig, J.; Vulule, J.; Irungu, L.; Walker, E.

2014-01-01

Clay pots were analyzed as devices for sampling the outdoor resting fraction of Anopheles gambiae Giles (Diptera: Culicidae) and other mosquito species in a rural, western Kenya. Clay pots (Anopheles gambiae resting pots, herein AgREPOTs), outdoor pit shelters, indoor pyrethrum spray collections (PSC), and Colombian curtain exit traps were compared in collections done biweekly for nine intervals from April to June 2005 in 20 housing compounds. Of 10,517 mosquitoes sampled, 4,668 An. gambiae s.l. were sampled in total of which 63% were An. gambiae s.s. (46% female) and 37% were An. arabiensis (66% female). The clay pots were useful and practical for sampling both sexes of An. gambiae s.l. Additionally, 617 An. funestus (58% female) and 5,232 Culex spp. (males and females together) were collected. Temporal changes in abundance of An. gambiae s.l. were similarly revealed by all four sampling methods, indicating that the clay pots could be used as devices to quantify variation in mosquito population density. Dispersion patterns of the different species and sexes fit well the negative binomial distribution, indicating that the mosquitoes were aggregated in distribution. Aside from providing a useful sampling tool, the AgREPOT also may be useful as a delivery vehicle for insecticides or pathogens to males and females that enter and rest in them. PMID:17294916

Anopheles gambiae complex (Diptera:Culicidae) near Bissau City, Guinea Bissau, West Africa.

PubMed

Fonseca, L F; Di Deco, M A; Carrara, G C; Dabo, I; Do Rosario, V; Petrarca, V

1996-11-01

Cytogenetic studies on mosquitoes collected inside bednets near Bissau City confirmed the presence of Anopheles melas Theobald and An. gambiae Giles sensu stricto, the latter species prevailing in rainy season samples (approximately 80% in average) and the former in dry season samples (> 90%). Seasonal and ecogeographical variations in the frequency of species and chromosomal inversions were analyzed. The analysis of An. gambiae sensu stricto confirmed the existence of the Bissau chromosomal form. The deficiency of heterokaryotypes in most samples indicated the possible coexistence of another chromosomal form not completely panmictic (i.e., randomly mating) with the Bissau form.

Evaluating preservation methods for identifying Anopheles gambiae s.s. and Anopheles arabiensis complex mosquitoes species using near infra-red spectroscopy

USDA-ARS?s Scientific Manuscript database

Near infra-red spectroscopy (NIRS) has been successfully used on fresh and RNAlater® preserved Anopheles gambiae complex to identify sibling species and age. No preservation methods other than RNAlater® have been tested to preserve mosquitoes for species identification using NIRS. However, RNAlater®...

Development of vegetable farming: a cause of the emergence of insecticide resistance in populations of Anopheles gambiae in urban areas of Benin

PubMed Central

Yadouleton, Anges William M; Asidi, Alex; Djouaka, Rousseau F; Braïma, James; Agossou, Christian D; Akogbeto, Martin C

2009-01-01

Background A fast development of urban agriculture has recently taken place in many areas in the Republic of Benin. This study aims to assess the rapid expansion of urban agriculture especially, its contribution to the emergence of insecticide resistance in populations of Anopheles gambiae. Methods The protocol was based on the collection of sociological data by interviewing vegetable farmers regarding various agricultural practices and the types of pesticides used. Bioassay tests were performed to assess the susceptibility of malaria vectors to various agricultural insecticides and biochemical analysis were done to characterize molecular status of population of An. gambiae. Results This research showed that: (1) The rapid development of urban agriculture is related to unemployment observed in cities, rural exodus and the search for a balanced diet by urban populations; (2) Urban agriculture increases the farmers' household income and their living standard; (3) At a molecular level, PCR revealed the presence of three sub-species of An. gambiae (An. gambiae s.s., Anopheles melas and Anopheles arabiensis) and two molecular forms (M and S). The kdr west mutation recorded in samples from the three sites and more specifically on the M forms seems to be one of the major resistance mechanisms found in An. gambiae from agricultural areas. Insecticide susceptibility tests conducted during this research revealed a clear pattern of resistance to permethrin (76% mortality rate at Parakou; 23.5% at Porto-Novo and 17% at Cotonou). Conclusion This study confirmed an increase activity of the vegetable farming in urban areas of Benin. This has led to the use of insecticide in an improper manner to control vegetable pests, thus exerting a huge selection pressure on mosquito larval population, which resulted to the emergence of insecticide resistance in malaria vectors. PMID:19442297

Spatial repellency screening in a high-throughput apparatus with Aedes aegypti and Anopheles gambiae

USDA-ARS?s Scientific Manuscript database

Spatial repellents are essential for personal protection against mosquitoes, such as Aedes aegypti and Anopheles gambiae, to reduce annoyance biting and transmission of mosquito-borne diseases. The number of safe and effective repellents, including DEET, picaridin, and IR3535, is limited and contin...

Geographic and ecologic distributions of the Anopheles gambiae complex predicted using a genetic algorithm.

PubMed

Levine, Rebecca S; Peterson, A Townsend; Benedict, Mark Q

2004-02-01

The distribution of the Anopheles gambiae complex of malaria vectors in Africa is uncertain due to under-sampling of vast regions. We use ecologic niche modeling to predict the potential distribution of three members of the complex (A. gambiae, A. arabiensis, and A. quadriannulatus) and demonstrate the statistical significance of the models. Predictions correspond well to previous estimates, but provide detail regarding spatial discontinuities in the distribution of A. gambiae s.s. that are consistent with population genetic studies. Our predictions also identify large areas of Africa where the presence of A. arabiensis is predicted, but few specimens have been obtained, suggesting under-sampling of the species. Finally, we project models developed from African distribution data for the late 1900s into the past and to South America to determine retrospectively whether the deadly 1929 introduction of A. gambiae sensu lato into Brazil was more likely that of A. gambiae sensu stricto or A. arabiensis.

Long non-coding RNA discovery across the genus anopheles reveals conserved secondary structures within and beyond the Gambiae complex.

PubMed

Jenkins, Adam M; Waterhouse, Robert M; Muskavitch, Marc A T

2015-04-23

Long non-coding RNAs (lncRNAs) have been defined as mRNA-like transcripts longer than 200 nucleotides that lack significant protein-coding potential, and many of them constitute scaffolds for ribonucleoprotein complexes with critical roles in epigenetic regulation. Various lncRNAs have been implicated in the modulation of chromatin structure, transcriptional and post-transcriptional gene regulation, and regulation of genomic stability in mammals, Caenorhabditis elegans, and Drosophila melanogaster. The purpose of this study is to identify the lncRNA landscape in the malaria vector An. gambiae and assess the evolutionary conservation of lncRNAs and their secondary structures across the Anopheles genus. Using deep RNA sequencing of multiple Anopheles gambiae life stages, we have identified 2,949 lncRNAs and more than 300 previously unannotated putative protein-coding genes. The lncRNAs exhibit differential expression profiles across life stages and adult genders. We find that across the genus Anopheles, lncRNAs display much lower sequence conservation than protein-coding genes. Additionally, we find that lncRNA secondary structure is highly conserved within the Gambiae complex, but diverges rapidly across the rest of the genus Anopheles. This study offers one of the first lncRNA secondary structure analyses in vector insects. Our description of lncRNAs in An. gambiae offers the most comprehensive genome-wide insights to date into lncRNAs in this vector mosquito, and defines a set of potential targets for the development of vector-based interventions that may further curb the human malaria burden in disease-endemic countries.

Analysis of near infrared spectra for age-grading of wild populations of Anopheles gambiae

USDA-ARS?s Scientific Manuscript database

A greater understanding of the age-structure of mosquito populations, especially malaria vectors such as Anopheles gambiae, is important for assessing the risk of infectious mosquitoes, and how vector control interventions may affect this structure. The use of near-infrared spectroscopy (NIRS) for a...

Spatial distribution and male mating success of Anopheles gambiae swarms

PubMed Central

2011-01-01

Background Anopheles gambiae mates in flight at particular mating sites over specific landmarks known as swarm markers. The swarms are composed of males; females typically approach a swarm, and leave in copula. This mating aggregation looks like a lek, but appears to lack the component of female choice. To investigate the possible mechanisms promoting the evolution of swarming in this mosquito species, we looked at the variation in mating success between swarms and discussed the factors that structure it in light of the three major lekking models, known as the female preference model, the hotspot model, and the hotshot model. Results We found substantial variation in swarm size and in mating success between swarms. A strong correlation between swarm size and mating success was observed, and consistent with the hotspot model of lek formation, the per capita mating success of individual males did not increase with swarm size. For the spatial distribution of swarms, our results revealed that some display sites were more attractive to both males and females and that females were more attracted to large swarms. While the swarm markers we recognize help us in localizing swarms, they did not account for the variation in swarm size or in the swarm mating success, suggesting that mosquitoes probably are attracted to these markers, but also perceive and respond to other aspects of the swarming site. Conclusions Characterizing the mating system of a species helps understand how this species has evolved and how selective pressures operate on male and female traits. The current study looked at male mating success of An. gambiae and discussed possible factors that account for its variation. We found that swarms of An. gambiae conform to the hotspot model of lek formation. But because swarms may lack the female choice component, we propose that the An. gambiae mating system is a lek-like system that incorporates characteristics pertaining to other mating systems such as scramble

Anopheles gambiae genome reannotation through synthesis of ab initio and comparative gene prediction algorithms

PubMed Central

Li, Jun; Riehle, Michelle M; Zhang, Yan; Xu, Jiannong; Oduol, Frederick; Gomez, Shawn M; Eiglmeier, Karin; Ueberheide, Beatrix M; Shabanowitz, Jeffrey; Hunt, Donald F; Ribeiro, José MC; Vernick, Kenneth D

2006-01-01

Background Complete genome annotation is a necessary tool as Anopheles gambiae researchers probe the biology of this potent malaria vector. Results We reannotate the A. gambiae genome by synthesizing comparative and ab initio sets of predicted coding sequences (CDSs) into a single set using an exon-gene-union algorithm followed by an open-reading-frame-selection algorithm. The reannotation predicts 20,970 CDSs supported by at least two lines of evidence, and it lowers the proportion of CDSs lacking start and/or stop codons to only approximately 4%. The reannotated CDS set includes a set of 4,681 novel CDSs not represented in the Ensembl annotation but with EST support, and another set of 4,031 Ensembl-supported genes that undergo major structural and, therefore, probably functional changes in the reannotated set. The quality and accuracy of the reannotation was assessed by comparison with end sequences from 20,249 full-length cDNA clones, and evaluation of mass spectrometry peptide hit rates from an A. gambiae shotgun proteomic dataset confirms that the reannotated CDSs offer a high quality protein database for proteomics. We provide a functional proteomics annotation, ReAnoXcel, obtained by analysis of the new CDSs through the AnoXcel pipeline, which allows functional comparisons of the CDS sets within the same bioinformatic platform. CDS data are available for download. Conclusion Comprehensive A. gambiae genome reannotation is achieved through a combination of comparative and ab initio gene prediction algorithms. PMID:16569258

Mapping potential Anopheles gambiae s.l. larval distribution using remotely sensed climatic and environmental variables in Baringo, Kenya.

PubMed

Amadi, J A; Ong'amo, G O; Olago, D O; Oriaso, S O; Nyamongo, I K; Estambale, B B A

2018-06-21

Anopheles gambiae s.l. (Diptera: Culicidae) is responsible for the transmission of the devastating Plasmodium falciparum (Haemosporida: Plasmodiidae) strain of malaria in Africa. This study investigated the relationship between climate and environmental conditions and An. gambiae s.l. larvae abundance and modelled the larval distribution of this species in Baringo County, Kenya. Mosquito larvae were collected using a 350-mL dipper and a pipette once per month from December 2015 to December 2016. A random forest algorithm was used to generate vegetation cover classes. A negative binomial regression was used to model the association between remotely sensed climate (rainfall and temperature) and environmental (vegetation cover, vegetation health, topographic wetness and slope) factors and An. gambiae s.l. for December 2015. Anopheles gambiae s.l. was significantly more frequent in the riverine zone (P < 0.05, r = 0.59) compared with the lowland zone. Rainfall (b = 6.22, P < 0.001), slope (b = - 4.81, P = 0.012) and vegetation health (b = - 5.60, P = 0.038) significantly influenced the distribution of An. gambiae s.l. larvae. High An. gambiae s.l. abundance was associated with cropland and wetland environments. Effective malaria control will require zone-specific interventions such as a focused dry season vector control strategy in the riverine zone. © 2018 The Royal Entomological Society.

Isolation and characterization of Y chromosome sequences from the African malaria mosquito Anopheles gambiae.

PubMed Central

Krzywinski, Jaroslaw; Nusskern, Deborah R; Kern, Marcia K; Besansky, Nora J

2004-01-01

The karyotype of the African malaria mosquito Anopheles gambiae contains two pairs of autosomes and a pair of sex chromosomes. The Y chromosome, constituting approximately 10% of the genome, remains virtually unexplored, despite the recent completion of the A. gambiae genome project. Here we report the identification and characterization of Y chromosome sequences of total length approaching 150 kb. We developed 11 Y-specific PCR markers that consistently yielded male-specific products in specimens from both laboratory colony and natural populations. The markers are characterized by low sequence polymorphism in samples collected across Africa and by presence in more than one copy on the Y. Screening of the A. gambiae BAC library using these markers allowed detection of 90 Y-linked BAC clones. Analysis of the BAC sequences and other Y-derived fragments showed massive accumulation of a few transposable elements. Nevertheless, more complex sequences are apparently present on the Y; these include portions of an approximately 48-kb-long unmapped AAAB01008227 scaffold from the whole genome shotgun assembly. Anopheles Y appears not to harbor any of the genes identified in Drosophila Y. However, experiments suggest that one of the ORFs from the AAAB01008227 scaffold represents a fragment of a gene with male-specific expression. PMID:15082548

First report of the presence of L1014S Knockdown-resistance mutation in Anopheles gambiae s.s and Anopheles coluzzii from Togo, West Africa.

PubMed

Djègbè, Innocent; Akoton, Romaric; Tchigossou, Genevieve; Ahadji-Dabla, Koffi Mensah; Atoyebi, Seun Michael; Adéoti, Razack; Zeukeng, Francis; Ketoh, Guillaume Koffivi; Djouaka, Rousseau

2018-01-01

Background: To optimize the success of insecticide-based malaria control intervention, knowledge of the distribution of Anopheles gambiae species and insecticide resistance mechanisms is necessary. This paper reported an updated data on pyrethroids/DDT resistance in the An. gambiae s.l population from Togo.  Methods: From December 2013 to April 2015, females of indoor-resting An. gambiae s.l were captured in three locations belonging to three different ecological zones. Resistance to DDT, permethrin and deltamethrin was screened in F1 progeny of collected mosquitoes using WHO susceptibility tests. The identification of species of An. gambiae complex and the detection of kdr and ace.1 R allele were carried out using DNA-based molecular techniques. Results: An. gambiae from Kovié and Nangbéto were highly resistant to DDT and permethrin with mortalities rate ranging from 0.83% to 1.58% for DDT and zero to 8.54% for permethrin. Mosquitoes collected in Nangbéto displayed 81.53% mortality with deltamethrin. An. coluzzii and An. gambiae s.s were found in sympatry in Nangbéto and Mango . The allelic frequency of L1014F was high, ranging from 66 to 100% in both An. coluzzii and An. gambiae s.s . For the first time we detected the L1014S allele in both An. coluzzii and An. gambiae s.s. from Togo at the frequency ranging from 5% to 13% in all the sites. The kdr N1575Y was present at various frequencies in both species ranging from 10% to 45%. Both An. gambiae s.s. and An. coluzzii shared the ace1 R mutation in all investigated sites with allelic frequency ranging from 4% to 16%. Conclusion: These results showed that multiple mutations are involved in insecticides resistance in An. gambiae populations from Togo including the kdr L1014F, L1014S, and N1575Y and ace.1 R G119S mutations.

First report of the presence of L1014S Knockdown-resistance mutation in Anopheles gambiae s.s and Anopheles coluzzii from Togo, West Africa

PubMed Central

Djègbè, Innocent; Akoton, Romaric; Tchigossou, Genevieve; Ahadji-Dabla, Koffi Mensah; Atoyebi, Seun Michael; Adéoti, Razack; Zeukeng, Francis; Ketoh, Guillaume Koffivi; Djouaka, Rousseau

2018-01-01

Background: To optimize the success of insecticide-based malaria control intervention, knowledge of the distribution of Anopheles gambiae species and insecticide resistance mechanisms is necessary. This paper reported an updated data on pyrethroids/DDT resistance in the An. gambiae s.l population from Togo.  Methods: From December 2013 to April 2015, females of indoor-resting An. gambiae s.l were captured in three locations belonging to three different ecological zones. Resistance to DDT, permethrin and deltamethrin was screened in F1 progeny of collected mosquitoes using WHO susceptibility tests. The identification of species of An. gambiae complex and the detection of kdr and ace.1 R allele were carried out using DNA-based molecular techniques. Results: An. gambiae from Kovié and Nangbéto were highly resistant to DDT and permethrin with mortalities rate ranging from 0.83% to 1.58% for DDT and zero to 8.54% for permethrin. Mosquitoes collected in Nangbéto displayed 81.53% mortality with deltamethrin. An. coluzzii and An. gambiae s.s were found in sympatry in Nangbéto and Mango . The allelic frequency of L1014F was high, ranging from 66 to 100% in both An. coluzzii and An. gambiae s.s. For the first time we detected the L1014S allele in both An. coluzzii and An. gambiae s.s. from Togo at the frequency ranging from 5% to 13% in all the sites. The kdr N1575Y was present at various frequencies in both species ranging from 10% to 45%. Both An. gambiae s.s. and An. coluzzii shared the ace1 R mutation in all investigated sites with allelic frequency ranging from 4% to 16%. Conclusion: These results showed that multiple mutations are involved in insecticides resistance in An. gambiae populations from Togo including the kdr L1014F, L1014S, and N1575Y and ace.1 R G119S mutations. PMID:29707654

The Seasonality and Ecology of the Anopheles gambiae complex (Dipetra: Culicidae) in Liberia Using Molecular Identification.

PubMed

Fahmy, N T; Villinski, J T; Bolay, F; Stoops, C A; Tageldin, R A; Fakoli, L; Okasha, O; Obenauer, P J; Diclaro, J W

2015-05-01

Members of the Anopheles gambiae sensu lato (Giles) complex define a group of seven morphologically indistinguishable species, including the principal malaria vectors in Sub-Saharan Africa. Members of this complex differ in behavior and ability to transmit malaria; hence, precise identification of member species is critical to monitoring and evaluating malaria threat levels. We collected mosquitoes from five counties in Liberia every other month from May 2011 until May 2012, using various trapping techniques. A. gambiae complex members were identified using molecular techniques based on differences in the ribosomal DNA (rDNA) region between species and the molecular forms (S and M) of A. gambiae sensu stricto (s.s) specimens. In total, 1,696 A. gambiae mosquitoes were collected and identified. DNA was extracted from legs of each specimen with species identification determined by multiplex polymerase chain reaction using specific primers. The molecular forms (M or S) of A. gambiae s.s were determined by restriction fragment length polymorphism. Bivariate and multivariate logistic regression models identified environmental variables associated with genomic differentiation. Our results indicate widespread occurrence of A. gambiae s.s., the principal malaria vector in the complex, although two Anopheles melas Theobald/A. merus Donitz mosquitoes were detected. We found 72.6, 25.5, and 1.9% of A. gambiae s.s specimens were S, M, and hybrid forms, respectively. Statistical analysis indicates that the S form was more likely to be found in rural areas during rainy seasons and indoor catchments. This information will enhance vector control efforts in Liberia. Published by Oxford University Press on behalf of Entomological Society of America 2015. This work is written by US Government employees and is in the public domain in the US.

Application of a reverse dot blot DNA-DNA hydridization method to quantify host-feeding tendencies of two sibling species in the Anopheles gambiae complex.

PubMed

Fritz, M L; Miller, J R; Bayoh, M N; Vulule, J M; Landgraf, J R; Walker, E D

2013-12-01

A DNA-DNA hybridization method, reverse dot blot analysis (RDBA), was used to identify Anopheles gambiae s.s. and Anopheles arabiensis (Diptera: Culicidae) hosts. Of 299 blood-fed and semi-gravid An. gambiae s.l. collected from Kisian, Kenya, 244 individuals were identifiable to species; of these, 69.5% were An. arabiensis and 29.5% were An. gambiae s.s. Host identifications with RDBA were comparable with those of conventional polymerase chain reaction (PCR) followed by direct sequencing of amplicons of the vertebrate mitochondrial cytochrome b gene. Of the 174 amplicon-producing samples used to compare these two methods, 147 were identifiable by direct sequencing and 139 of these were identifiable by RDBA. Anopheles arabiensis bloodmeals were mostly (94.6%) bovine in origin, whereas An. gambiae s.s. fed upon humans more than 91.8% of the time. Tests by RDBA detected that two of 112 An. arabiensis contained blood from more than one host species, whereas PCR and direct sequencing did not. Recent use of insecticide-treated bednets in Kisian is likely to have caused the shift in the dominant vector species from An. gambiae s.s. to An. arabiensis. Reverse dot blot analysis provides an opportunity to study changes in host-feeding by members of the An. gambiae complex in response to the broadening distribution of vector control measures targeting host-selection behaviours. © 2013 The Royal Entomological Society.

Molecular karyotyping of the 2La inversion in Anopheles gambiae.

PubMed

White, Bradley J; Santolamazza, Federica; Kamau, Luna; Pombi, Marco; Grushko, Olga; Mouline, Karine; Brengues, Cecile; Guelbeogo, Wamdaogo; Coulibaly, Mamadou; Kayondo, Jonathan K; Sharakhov, Igor; Simard, Frederic; Petrarca, Vincenzo; Della Torre, Alessandra; Besansky, Nora J

2007-02-01

The African malaria vector Anopheles gambiae is polymorphic for alternative arrangements on the left arm of chromosome 2 (2La and 2L+(a)) that are non-randomly distributed with respect to degree of aridity. Detailed studies on the ecological role of inversion 2La have been hindered by the technical demands of traditional karyotype analysis and by sex- and stage-specific limitations on the availability of polytene chromosomes favorable for analysis. Recent molecular characterization of both inversion breakpoints presented the opportunity to develop a polymerase chain reaction (PCR)-based method for karyotype analysis. Here we report the development of this molecular diagnostic assay and the results of extensive field validation. When tested on 765 An. gambiae specimens sampled across Africa, the molecular approach compared favorably with traditional cytologic methods, correctly scoring > 94% of these specimens. By providing ready access to the 2La karyotype, this tool lays groundwork for future studies of the ecological genomics of this medically important species.

Seasonal climate effects anemotaxis in newly emerged adult anopheles gambiae giles in Mali, West Africa

USDA-ARS?s Scientific Manuscript database

The direction and magnitude of movement of the malaria vector Anopheles gambiae Giles has been of great interest to medical entomologists for over 70 years. This direction of movement is likely to be affected by many factors, from environmental conditions and stage of life history of the mosquito to...

Sugar-fermenting yeast as an organic source of carbon dioxide to attract the malaria mosquito Anopheles gambiae

PubMed Central

2010-01-01

Background Carbon dioxide (CO2) plays an important role in the host-seeking process of opportunistic, zoophilic and anthropophilic mosquito species and is, therefore, commonly added to mosquito sampling tools. The African malaria vector Anopheles gambiae sensu stricto is attracted to human volatiles augmented by CO2. This study investigated whether CO2, usually supplied from gas cylinders acquired from commercial industry, could be replaced by CO2 derived from fermenting yeast (yeast-produced CO2). Methods Trapping experiments were conducted in the laboratory, semi-field and field, with An. gambiae s.s. as the target species. MM-X traps were baited with volatiles produced by mixtures of yeast, sugar and water, prepared in 1.5, 5 or 25 L bottles. Catches were compared with traps baited with industrial CO2. The additional effect of human odours was also examined. In the laboratory and semi-field facility dual-choice experiments were conducted. The effect of traps baited with yeast-produced CO2 on the number of mosquitoes entering an African house was studied in the MalariaSphere. Carbon dioxide baited traps, placed outside human dwellings, were also tested in an African village setting. The laboratory and semi-field data were analysed by a χ2-test, the field data by GLM. In addition, CO2 concentrations produced by yeast-sugar solutions were measured over time. Results Traps baited with yeast-produced CO2 caught significantly more mosquitoes than unbaited traps (up to 34 h post mixing the ingredients) and also significantly more than traps baited with industrial CO2, both in the laboratory and semi-field. Adding yeast-produced CO2 to traps baited with human odour significantly increased trap catches. In the MalariaSphere, outdoor traps baited with yeast-produced or industrial CO2 + human odour reduced house entry of mosquitoes with a human host sleeping under a bed net indoors. Anopheles gambiae s.s. was not caught during the field trials. However, traps baited with

Evidence of carbamate resistance in urban populations of Anopheles gambiae s.s. mosquitoes resistant to DDT and deltamethrin insecticides in Lagos, South-Western Nigeria

PubMed Central

2012-01-01

Background Resistance monitoring is essential in ensuring the success of insecticide based vector control programmes. This study was carried out to assess the susceptibility status of urban populations of Anopheles gambiae to carbamate insecticide being considered for vector control in mosquito populations previously reported to be resistant to DDT and permethrin. Methods Two – three day old adult female Anopheles mosquitoes reared from larval collections in 11 study sites from Local Government Areas of Lagos were exposed to test papers impregnated with DDT 4%, deltamethrin 0.05% and propoxur 0.1% insecticides. Additional tests were carried out to determine the susceptibility status of the Anopheles gambiae population to bendiocarb insecticide. Members of the A. gambiae complex, the molecular forms, were identified by PCR assays. The involvement of metabolic enzymes in carbamate resistance was assessed using Piperonyl butoxide (PBO) synergist assays. The presence of kdr-w/e and ace-1R point mutations responsible for DDT-pyrethroid and carbamate resistance mechanisms was also investigated by PCR. Results Propoxur resistance was found in 10 out of the 11 study sites. Resistance to three classes of insecticides was observed in five urban localities. Mortality rates in mosquitoes exposed to deltamethrin and propoxur did not show any significant difference (P > 0.05) but was significantly higher (P < 0.05) in populations exposed to DDT. All mosquitoes tested were identified as A. gambiae s.s (M form). The kdr -w point mutation at allelic frequencies between 45%-77% was identified as one of the resistant mechanisms responsible for DDT and pyrethroid resistance. Ace-1R point mutation was absent in the carbamate resistant population. However, the possible involvement of metabolic resistance was confirmed by synergistic assays conducted. Conclusion Evidence of carbamate resistance in A. gambiae populations already harbouring resistance to DDT and permethrin is a

Transcription profiling of a recently colonised pyrethroid resistant Anopheles gambiae strain from Ghana

PubMed Central

Müller, Pie; Donnelly, Martin J; Ranson, Hilary

2007-01-01

Background Mosquito resistance to the pyrethroid insecticides used to treat bednets threatens the sustainability of malaria control in sub-Saharan Africa. While the impact of target site insensitivity alleles is being widely discussed the implications of insecticide detoxification – though equally important – remains elusive. The successful development of new tools for malaria intervention and management requires a comprehensive understanding of insecticide resistance, including metabolic resistance mechanisms. Although three enzyme families (cytochrome P450s, glutathione S-transferases and carboxylesterases) have been widely associated with insecticide detoxification the role of individual enzymes is largely unknown. Results Here, constitutive expression patterns of genes putatively involved in conferring pyrethroid resistance was investigated in a recently colonised pyrethroid resistant Anopheles gambiae strain from Odumasy, Southern Ghana. RNA from the resistant strain and a standard laboratory susceptible strain, of both sexes was extracted, reverse transcribed and labelled with either Cy3- or Cy5-dye. Labelled cDNA was co-hybridised to the detox chip, a custom-made microarray containing over 230 A. gambiae gene fragments predominantly from enzyme families associated with insecticide resistance. After hybridisation, Cy3- and Cy5-signal intensities were measured and compared gene by gene. In both females and males of the resistant strain the cytochrome P450s CYP6Z2 and CYP6M2 are highly over-expressed along with a member of the superoxide dismutase (SOD) gene family. Conclusion These genes differ from those found up-regulated in East African strains of pyrethroid resistant A. gambiae and constitute a novel set of candidate genes implicated in insecticide detoxification. These data suggest that metabolic resistance may have multiple origins in A. gambiae, which has strong implications for the management of resistance. PMID:17261191

Pyrethroid, DDT and malathion resistance in the malaria vector Anopheles gambiae from the Democratic Republic of Congo.

PubMed

Basilua Kanza, Jean Pierre; El Fahime, Elmostafa; Alaoui, Sanaa; Essassi, El Mokhtar; Brooke, Basil; Nkebolo Malafu, André; Watsenga Tezzo, Francis

2013-01-01

Malaria remains the most important parasitic disease in sub-Saharan Africa. We investigated the extent of resistance in the malaria vector Anopheles gambiae from the Democratic Republic of Congo (DRC) to three classes of insecticide approved by WHO for indoor residual spraying. Standard WHO bioassays were performed on adult Anopheles mosquitoes reared in the laboratory from larvae collected from different sites. Molecular techniques were used for species identification and to identify knockdown resistance (kdr) and acetylcholinesterase (ace-1(R)) mutations in individual mosquitoes. Only A. gambiae s.s., the nominal member of the A. gambiae species complex, was found. Bioassays showed phenotypic resistance to the main insecticides used in the region, notably pyrethroids (deltamethrin, permethrin, lambda-cyhalothrin), an organochlorine (DDT) and an organophosphate (malathion). The L1014F kdr allele, often associated with resistance to pyrethroids and DDT, was detected in samples from all collection sites at varying frequencies. No ace-1(R) resistance alleles (associated with organophosphate and carbamate resistance) were detected. These data can be used to inform a resistance management strategy that requires comprehensive information concerning malaria vector species composition in the areas of interest, and their susceptibility to the insecticides proposed for their control.

A test of the chromosomal theory of ecotypic speciation in Anopheles gambiae

PubMed Central

Manoukis, Nicholas C.; Powell, Jeffrey R.; Touré, Mahamoudou B.; Sacko, Adama; Edillo, Frances E.; Coulibaly, Mamadou B.; Traoré, Sekou F.; Taylor, Charles E.; Besansky, Nora J.

2008-01-01

The role of chromosomal inversions in speciation has long been of interest to evolutionists. Recent quantitative modeling has stimulated reconsideration of previous conceptual models for chromosomal speciation. Anopheles gambiae, the most important vector of human malaria, carries abundant chromosomal inversion polymorphism nonrandomly associated with ecotypes that mate assortatively. Here, we consider the potential role of paracentric inversions in promoting speciation in A. gambiae via “ecotypification,” a term that refers to differentiation arising from local adaptation. In particular, we focus on the Bamako form, an ecotype characterized by low inversion polymorphism and fixation of an inversion, 2Rj, that is very rare or absent in all other forms of A. gambiae. The Bamako form has a restricted distribution by the upper Niger River and its tributaries that is associated with a distinctive type of larval habitat, laterite rock pools, hypothesized to be its optimal breeding site. We first present computer simulations to investigate whether the population dynamics of A. gambiae are consistent with chromosomal speciation by ecotypification. The models are parameterized using field observations on the various forms of A. gambiae that exist in Mali, West Africa. We then report on the distribution of larvae of this species collected from rock pools and more characteristic breeding sites nearby. Both the simulations and field observations support the thesis that speciation by ecotypification is occurring, or has occurred, prompting consideration of Bamako as an independent species. PMID:18287019

Rapid discrimination between Anopheles gambiae s.s. and Anopheles arabiensis by High-Resolution Melt (HRM) analysis.

PubMed

Zianni, Michael R; Nikbakhtzadeh, Mahmood R; Jackson, Bryan T; Panescu, Jenny; Foster, Woodbridge A

2013-04-01

There is a need for more cost-effective options to more accurately discriminate among members of the Anopheles gambiae complex, particularly An. gambiae and Anopheles arabiensis. These species are morphologically indistinguishable in the adult stage, have overlapping distributions, but are behaviorally and ecologically different, yet both are efficient vectors of malaria in equatorial Africa. The method described here, High-Resolution Melt (HRM) analysis, takes advantage of minute differences in DNA melting characteristics, depending on the number of incongruent single nucleotide polymorphisms in an intragenic spacer region of the X-chromosome-based ribosomal DNA. The two species in question differ by an average of 13 single-nucleotide polymorphisms giving widely divergent melting curves. A real-time PCR system, Bio-Rad CFX96, was used in combination with a dsDNA-specific dye, EvaGreen, to detect and measure the melting properties of the amplicon generated from leg-extracted DNA of selected mosquitoes. Results with seven individuals from pure colonies of known species, as well as 10 field-captured individuals unambiguously identified by DNA sequencing, demonstrated that the method provided a high level of accuracy. The method was used to identify 86 field mosquitoes through the assignment of each to the two common clusters with a high degree of certainty. Each cluster was defined by individuals from pure colonies. HRM analysis is simpler to use than most other methods and provides comparable or more accurate discrimination between the two sibling species but requires a specialized melt-analysis instrument and software.

Rapid Discrimination between Anopheles gambiae s.s. and Anopheles arabiensis by High-Resolution Melt (HRM) Analysis

PubMed Central

Zianni, Michael R.; Nikbakhtzadeh, Mahmood R.; Jackson, Bryan T.; Panescu, Jenny; Foster, Woodbridge A.

2013-01-01

There is a need for more cost-effective options to more accurately discriminate among members of the Anopheles gambiae complex, particularly An. gambiae and Anopheles arabiensis. These species are morphologically indistinguishable in the adult stage, have overlapping distributions, but are behaviorally and ecologically different, yet both are efficient vectors of malaria in equatorial Africa. The method described here, High-Resolution Melt (HRM) analysis, takes advantage of minute differences in DNA melting characteristics, depending on the number of incongruent single nucleotide polymorphisms in an intragenic spacer region of the X-chromosome-based ribosomal DNA. The two species in question differ by an average of 13 single-nucleotide polymorphisms giving widely divergent melting curves. A real-time PCR system, Bio-Rad CFX96, was used in combination with a dsDNA-specific dye, EvaGreen, to detect and measure the melting properties of the amplicon generated from leg-extracted DNA of selected mosquitoes. Results with seven individuals from pure colonies of known species, as well as 10 field-captured individuals unambiguously identified by DNA sequencing, demonstrated that the method provided a high level of accuracy. The method was used to identify 86 field mosquitoes through the assignment of each to the two common clusters with a high degree of certainty. Each cluster was defined by individuals from pure colonies. HRM analysis is simpler to use than most other methods and provides comparable or more accurate discrimination between the two sibling species but requires a specialized melt-analysis instrument and software. PMID:23543777

Larvicidal effects of a neem (Azadirachta indica) oil formulation on the malaria vector Anopheles gambiae

PubMed Central

Okumu, Fredros O; Knols, Bart GJ; Fillinger, Ulrike

2007-01-01

Background Larviciding is a key strategy used in many vector control programmes around the world. Costs could be reduced if larvicides could be manufactured locally. The potential of natural products as larvicides against the main African malaria vector, Anopheles gambiae s.s was evaluated. Methods To assess the larvicidal efficacy of a neem (Azadirachta indica) oil formulation (azadirachtin content of 0.03% w/v) on An. gambiae s.s., larvae were exposed as third and fourth instars to a normal diet supplemented with the neem oil formulations in different concentrations. A control group of larvae was exposed to a corn oil formulation in similar concentrations. Results Neem oil had an LC50 value of 11 ppm after 8 days, which was nearly five times more toxic than the corn oil formulation. Adult emergence was inhibited by 50% at a concentration of 6 ppm. Significant reductions on growth indices and pupation, besides prolonged larval periods, were observed at neem oil concentrations above 8 ppm. The corn oil formulation, in contrast, produced no growth disruption within the tested range of concentrations. Conclusion Neem oil has good larvicidal properties for An. gambiae s.s. and suppresses successful adult emergence at very low concentrations. Considering the wide distribution and availability of this tree and its products along the East African coast, this may prove a readily available and cheap alternative to conventional larvicides. PMID:17519000

Distribution of genetic diversity in relation to chromosomal inversions in the malaria mosquito Anopheles gambiae.

PubMed

Mathiopoulos, K D; Lanzaro, G C

1995-06-01

The epidemiology of malaria in Africa is complicated by the fact that its principal vector, the mosquito Anopheles gambiae, constitutes a complex of six sibling species. Each species is characterized by a unique array of paracentric inversions, as deduced by karyotypic analysis. In addition, most of the species carry a number of polymorphic inversions. In order to develop an understanding of the evolutionary histories of different parts of the genome, we compared the genetic variation of areas inside and outside inversions in two distinct inversion karyotypes of A. gambiae. Thirty-five cDNA clones were mapped on the five arms of the A. gambiae chromosomes with divisional probes. Sixteen of these clones, localized both inside and outside inversions of chromosome 2, were used as probes in order to determine the nucleotide diversity of different parts of the genome in the two inversion karyotypes. We observed that the sequence diversity inside the inversion is more than three-fold lower than in areas outside the inversion and that the degree of divergence increases gradually at loci at increasing distance from the inversion. To interpret the data we present a selectionist and a stochastic model, both of which point to a relatively recent origin of the studied inversion and may suggest differences between the evolutionary history of inversions in Anopheles and Drosophila species.

Pyrethroids and DDT tolerance of Anopheles gambiae s.l. from Sengerema District, an area of intensive pesticide usage in north-western Tanzania.

PubMed

Philbert, Anitha; Lyantagaye, Sylvester Leonard; Pradel, Gabriele; Ngwa, Che Julius; Nkwengulila, Gamba

2017-04-01

To assess the susceptibility status of malaria vectors to pyrethroids and dichlorodiphenyltrichloroethane (DDT), characterise the mechanisms underlying resistance and evaluate the role of agro-chemical use in resistance selection among malaria vectors in Sengerema agro-ecosystem zone, Tanzania. Mosquito larvae were collected from farms and reared to obtain adults. The susceptibility status of An. gambiae s.l. was assessed using WHO bioassay tests to permethrin, deltamethrin, lambdacyhalothrin, etofenprox, cyfluthrin and DDT. Resistant specimens were screened for knock-down resistance gene (kdr), followed by sequencing both Western and Eastern African variants. A gas chromatography-mass spectrophotometer (GC-MS) was used to determine pesticide residues in soil and sediments from mosquitoes' breeding habitats. Anopheles gambiae s.l. was resistant to all the insecticides tested. The population of Anopheles gambiae s.l was composed of Anopheles arabiensis by 91%. The East African kdr (L1014S) allele was found in 13 of 305 specimens that survived insecticide exposure, with an allele frequency from 0.9% to 50%. DDTs residues were found in soils at a concentration up to 9.90 ng/g (dry weight). The observed high resistance levels of An. gambiae s.l., the detection of kdr mutations and pesticide residues in mosquito breeding habitats demonstrate vector resistance mediated by pesticide usage. An integrated intervention through collaboration of agricultural, livestock and vector control units is vital. © 2017 John Wiley & Sons Ltd.

Inversions and Gene Order Shuffling in Anopheles gambiae and A. funestus

NASA Astrophysics Data System (ADS)

Sharakhov, Igor V.; Serazin, Andrew C.; Grushko, Olga G.; Dana, Ali; Lobo, Neil; Hillenmeyer, Maureen E.; Westerman, Richard; Romero-Severson, Jeanne; Costantini, Carlo; Sagnon, N'Fale; Collins, Frank H.; Besansky, Nora J.

2002-10-01

In tropical Africa, Anopheles funestus is one of the three most important malaria vectors. We physically mapped 157 A. funestus complementary DNAs (cDNAs) to the polytene chromosomes of this species. Sequences of the cDNAs were mapped in silico to the A. gambiae genome as part of a comparative genomic study of synteny, gene order, and sequence conservation between A. funestus and A. gambiae. These species are in the same subgenus and diverged about as recently as humans and chimpanzees. Despite nearly perfect preservation of synteny, we found substantial shuffling of gene order along corresponding chromosome arms. Since the divergence of these species, at least 70 chromosomal inversions have been fixed, the highest rate of rearrangement of any eukaryote studied to date. The high incidence of paracentric inversions and limited colinearity suggests that locating genes in one anopheline species based on gene order in another may be limited to closely related taxa.

[Cytogenetic and biometric observations on members of the Anopheles gambiae complex in Mozambique].

PubMed

Petrarca, V; Carrara, G C; Di Deco, M A; Petrangeli, G

1984-12-01

Four species of the Anopheles gambiae complex were identified in Mozambique (East Africa) by chromosomal analysis. They were An. merus, An. gambiae s.s., An. arabiensis and An. quadriannulatus. An. merus was observed in coastal zones as well as in inner areas where the rivers are tidal and brackish and/or the soil is salty. An. gambiae s.s. is present in the central-northern regions (north of Save river) from the coast to the western mountains. On the coast it is often sympatric with An. merus. It is apparently absent south of Save river. An. arabiensis was observed in samples from the north-western hilly and mountainous areas, sympatrically with An. gambiae s.s., as well as south of Save river where often it is sympatric with An. merus. Only one specimen of An. quadriannulatus was observed. It was from a small sample collected feeding on bovid in a southern locality (Bela Vista-Maputo area). No inversion polymorphism was observed in the 446 An. merus identified. A quite low degree of inversion polymorphism was shown by both An. gambiae s.s. and An. arabiensis, involving 2Rb, 2La and 2Rb inversion systems respectively. By morphological analysis of cytogenetically identified samples of three species of the complex, number of sensilla coeloconica and palpal ratio were confirmed to be useful to distinguish An. merus from An. gambiae s.s./An. arabiensis. The overlapping areas between brackish and freshwater species become smaller when both characters are considered together, 1.5% and 3.5% being the probabilities of confusing An. merus with An. gambiae s.s. and An. arabiensis respectively.

Discovery and characterization of two Nimrod superfamily members in Anopheles gambiae.

PubMed

Midega, Janet; Blight, Joshua; Lombardo, Fabrizio; Povelones, Michael; Kafatos, Fotis; Christophides, George K

2013-12-01

Anti-bacterial proteins in mosquitoes are known to play an important modulatory role on immune responses to infections with human pathogens including malaria parasites. In this study we characterized two members of the Anopheles gambiae Nimrod superfamily, namely AgNimB2 and AgEater. We confirm that current annotation of the An. gambiae genome incorrectly identifies AgNimB2 and AgEater as a single gene, AGAP009762. Through in silico and experimental approaches, it has been shown that AgNimB2 is a secreted protein that mediates phagocytosis of Staphylococcus aureus but not of Escherichia coli bacteria. We also reveal that this function does not involve a direct interaction of AgNimB2 with S. aureus. Therefore, AgNimB2 may act downstream of complement-like pathway activation, first requiring bacterial opsonization. In addition, it has been shown that AgNimB2 has an anti-Plasmodium effect. Conversely, AgEater is a membrane-bound protein that either functions redundantly or is dispensable for phagocytosis of E. coli or S. aureus. Our study provides insights into the role of members of the complex Nimrod superfamily in An. gambiae, the most important African vector of human malaria.

Discovery and characterization of two Nimrod superfamily members in Anopheles gambiae

PubMed Central

Midega, Janet; Blight, Joshua; Lombardo, Fabrizio; Povelones, Michael; Kafatos, Fotis; Christophides, George K

2013-01-01

Anti-bacterial proteins in mosquitoes are known to play an important modulatory role on immune responses to infections with human pathogens including malaria parasites. In this study we characterized two members of the Anopheles gambiae Nimrod superfamily, namely AgNimB2 and AgEater. We confirm that current annotation of the An. gambiae genome incorrectly identifies AgNimB2 and AgEater as a single gene, AGAP009762. Through in silico and experimental approaches, it has been shown that AgNimB2 is a secreted protein that mediates phagocytosis of Staphylococcus aureus but not of Escherichia coli bacteria. We also reveal that this function does not involve a direct interaction of AgNimB2 with S. aureus. Therefore, AgNimB2 may act downstream of complement-like pathway activation, first requiring bacterial opsonization. In addition, it has been shown that AgNimB2 has an anti-Plasmodium effect. Conversely, AgEater is a membrane-bound protein that either functions redundantly or is dispensable for phagocytosis of E. coli or S. aureus. Our study provides insights into the role of members of the complex Nimrod superfamily in An. gambiae, the most important African vector of human malaria. PMID:24428830

Daily rhythms in antennal protein and olfactory sensitivity in the malaria mosquito Anopheles gambiae

PubMed Central

Rund, Samuel S. C.; Bonar, Nicolle A.; Champion, Matthew M.; Ghazi, John P.; Houk, Cameron M.; Leming, Matthew T.; Syed, Zainulabeuddin; Duffield, Giles E.

2013-01-01

We recently characterized 24-hr daily rhythmic patterns of gene expression in Anopheles gambiae mosquitoes. These include numerous odorant binding proteins (OBPs), soluble odorant carrying proteins enriched in olfactory organs. Here we demonstrate that multiple rhythmically expressed genes including OBPs and takeout proteins, involved in regulating blood feeding behavior, have corresponding rhythmic protein levels as measured by quantitative proteomics. This includes AgamOBP1, previously shown as important to An. gambiae odorant sensing. Further, electrophysiological investigations demonstrate time-of-day specific differences in olfactory sensitivity of antennae to major host-derived odorants. The pre-dusk/dusk peaks in OBPs and takeout gene expression correspond with peak protein abundance at night, and in turn coincide with the time of increased olfactory sensitivity to odorants requiring OBPs and times of increased blood-feeding behavior. This suggests an important role for OBPs in modulating temporal changes in odorant sensitivity, enabling the olfactory system to coordinate with the circadian niche of An. gambiae. PMID:23986098

Exposure to disinfectants (soap or hydrogen peroxide) increases tolerance to permethrin in Anopheles gambiae populations from the city of Yaoundé, Cameroon

PubMed Central

2014-01-01

Background The rapid expansion of insecticide resistance is limiting the efficiency of malaria vector control interventions. However, current knowledge of factors inducing pyrethroid resistance remains incomplete. In the present study, the role of selection at the larval stage by disinfectants, such as soap and hydrogen peroxide (H2O2), on adult mosquito resistance to permethrin was investigated. Methods Field Anopheles gambiae sensu lato larvae, were exposed to variable concentrations of soap and H2O2. Larvae surviving to acute toxicity assays after 24 hours were reared to the adult stage and exposed to permethrin. The susceptibility level of adults was compared to the untreated control group. The effect of soap or hydrogen peroxide selection on the length of larval development and emergence rate was assessed. Result Larval bioassays analysis showed a more acute effect of hydrogen peroxide on mosquito larvae compared to soap. The regression lines describing the dose mortality profile showed higher mean and variance to hydrogen peroxide than to soap. The duration of larval development (<5 days) and adults emergence rates (1 to 77%) were shorter and lower compare to control. Anopheles gambiae s.l. larvae surviving to selection with either soap or hydrogen peroxide or both, produced adults who were up to eight-times more resistant to permethrin than mosquitoes from the untreated control group. Conclusion The present study shows that selective pressure exerted by non-insecticidal compounds such as soap and hydrogen peroxide affect An. gambiae s.l. tolerance to pyrethroids. This requires further studies with regard to the adaptation of An. gambiae s.l. to polluted habitats across sub-Saharan Africa cities. PMID:25086741

Susceptibility of Anopheles gambiae to insecticides used for malaria vector control in Rwanda.

PubMed

Hakizimana, Emmanuel; Karema, Corine; Munyakanage, Dunia; Iranzi, Gad; Githure, John; Tongren, Jon Eric; Takken, Willem; Binagwaho, Agnes; Koenraadt, Constantianus J M

2016-12-01

The widespread emergence of resistance to pyrethroids is a major threat to the gains made in malaria control. To monitor the presence and possible emergence of resistance against a variety of insecticides used for malaria control in Rwanda, nationwide insecticide resistance surveys were conducted in 2011 and 2013. Larvae of Anopheles gambiae sensu lato mosquitoes were collected in 12 sentinel sites throughout Rwanda. These were reared to adults and analysed for knock-down and mortality using WHO insecticide test papers with standard diagnostic doses of the recommended insecticides. A sub-sample of tested specimens was analysed for the presence of knockdown resistance (kdr) mutations. A total of 14,311 mosquitoes were tested and from a sample of 1406 specimens, 1165 (82.9%) were identified as Anopheles arabiensis and 241 (17.1%) as Anopheles gambiae sensu stricto. Mortality results indicated a significant increase in resistance to lambda-cyhalothrin from 2011 to 2013 in 83% of the sites, permethrin in 25% of the sites, deltamethrin in 25% of the sites and DDT in 50% of the sites. Mosquitoes from 83% of the sites showed full susceptibility to bendiocarb and 17% of sites were suspected to harbour resistance that requires further confirmation. No resistance was observed to fenitrothion in all study sites during the entire survey. The kdr genotype results in An. gambiae s.s. showed that 67 (50%) possessed susceptibility (SS) alleles, while 35 (26.1%) and 32 (23.9%) mosquitoes had heterozygous (RS) and homozygous (RR) alleles, respectively. Of the 591 An. arabiensis genotyped, 425 (71.9%) possessed homozygous (SS) alleles while 158 (26.7%) and 8 (1.4%) had heterozygous (RS) and homozygous (RR) alleles, respectively. Metabolic resistance involving oxidase enzymes was also detected using the synergist PBO. This is the first nationwide study of insecticide resistance in malaria vectors in Rwanda. It shows the gradual increase of insecticide resistance to pyrethroids (lambda

Proteomics reveals novel components of the Anopheles gambiae eggshell

PubMed Central

Amenya, Dolphine A.; Chou, Wayne; Li, Jianyong; Yan, Guiyun; Gershon, Paul D.; James, Anthony A.; Marinotti, Osvaldo

2010-01-01

While genome and transcriptome sequencing has revealed a large number and diversity of Anopheles gambiae predicted proteins, identifying their functions and biosynthetic pathways remains challenging. Applied mass spectrometry based proteomics in conjunction with mosquito genome and transcriptome databases were used to identify 44 proteins as putative components of the eggshell. Among the identified molecules are two vitelline membrane proteins and a group of seven putative chorion proteins. Enzymes with peroxidase, laccase and phenoloxidase activities, likely involved in cross-linking reactions that stabilize the eggshell structure, also were identified. Seven odorant binding proteins were found in association with the mosquito eggshell, although their role has yet to be demonstrated. This analysis fills a considerable gap of knowledge about proteins that build the eggshell of anopheline mosquitoes. PMID:20433845

Behavioral response of the malaria mosquito, Anopheles gambiae, to human sweat inoculated with axilla bacteria and to volatiles composing human axillary odor.

PubMed

Frei, Jérôme; Kröber, Thomas; Troccaz, Myriam; Starkenmann, Christian; Guerin, Patrick M

2017-02-01

The responses of Anopheles gambiae Giles sensu stricto (Diptera: Culicidae) to odors from male and female axillary sweat incubated with human axilla bacteria were recorded in a dual-choice olfactometer. Staphylococcus epidermidis was selected for its low odor-producing pattern, Corynebacterium jeikeium for its strong Nα-acylglutamine aminoacylase activity liberating carboxylic acids including (R)/(S)-3-hydroxy-3-methylhexanoic acid (HMHA) and Staphylococcus haemolyticus for its capacity to liberate sulfur-containing compounds including (R/S)-3-methyl-3-sulfanylhexan-1-ol (MSH). Anopheles gambiae behavioral responses were evaluated under (i) its responsiveness to take off and undertake sustained upwind flight and (ii) its discriminating capacity between the two olfactometer arms bearing a test odor in either one or both arms. Experiments were conducted in the presence of carbon dioxide pulses as a behavioral sensitizer. Anopheles gambiae clearly discriminated for the olfactometer arm conveying odor generated by incubating any of the three bacteria species with either male or female sweat. Whereas An. gambiae did not discriminate between male and female sterile sweat samples in the olfactometer, the mosquito consistently showed a preference for male sweat over female sweat incubated with the same bacterium, independent of the species used as inoculum. Sweat incubated with C. jeikeium rendered mosquitoes particularly responsive and this substrate elicited the strongest preference for male over female sweat. Tested on their own, neither HMHA nor MSH elicited a clear discriminating response but did affect mosquito responsiveness. These findings serve as a basis for further research on the odor-mediated anthropophilic host-seeking behavior of An. gambiae. © The Author 2016. Published by Oxford University Press. All rights reserved. For permissions, please e-mail: journals.permissions@oup.com.

Multiple insecticide resistance mechanisms in Anopheles gambiae s.l. populations from Cameroon, Central Africa.

PubMed

Nwane, Philippe; Etang, Josiane; Chouaїbou, Mouhamadou; Toto, Jean Claude; Koffi, Alphonsine; Mimpfoundi, Rémy; Simard, Frédéric

2013-02-22

Increasing incidence of DDT and pyrethroid resistance in Anopheles mosquitoes is seen as a limiting factor for malaria vector control. The current study aimed at an in-depth characterization of An. gambiae s.l. resistance to insecticides in Cameroon, in order to guide malaria vector control interventions. Anopheles gambiae s.l. mosquitoes were collected as larvae and pupae from six localities spread throughout the four main biogeographical domains of Cameroon and reared to adults in insectaries. Standard WHO insecticide susceptibility tests were carried out with 4% DDT, 0.75% permethrin and 0.05% deltamethrin. Mortality rates and knockdown times (kdt50 and kdt95) were determined and the effect of pre-exposure to the synergists DEF, DEM and PBO was assessed. Tested mosquitoes were identified to species and molecular forms (M or S) using PCR-RFLP. The hot ligation method was used to depict kdr mutations and biochemical assays were conducted to assess detoxifying enzyme activities. The An. arabiensis population from Pitoa was fully susceptible to DDT and permethrin (mortality rates>98%) and showed reduced susceptibility to deltamethrin. Resistance to DDT was widespread in An. gambiae s.s. populations and heterogeneous levels of susceptibility to permethrin and deltamethrin were observed. In many cases, prior exposure to synergists partially restored insecticide knockdown effect and increased mortality rates, suggesting a role of detoxifying enzymes in increasing mosquito survival upon challenge by pyrethroids and, to a lower extent DDT. The distribution of kdr alleles suggested a major role of kdr-based resistance in the S form of An. gambiae. In biochemical tests, all but one mosquito population overexpressed P450 activity, whereas baseline GST activity was low and similar in all field mosquito populations and in the control. In Cameroon, multiple resistance mechanisms segregate in the S form of An. gambiae resulting in heterogeneous resistance profiles, whereas in

Chemical composition and insecticidal activity of plant essential oils from Benin against Anopheles gambiae (Giles)

PubMed Central

2013-01-01

Background Insecticide resistance in sub-Saharan Africa and especially in Benin is a major public health issue hindering the control of the malaria vectors. Each Anopheles species has developed a resistance to one or several classes of the insecticides currently in use in the field. Therefore, it is urgent to find alternative compounds to conquer the vector. In this study, the efficacies of essential oils of nine plant species, which are traditionally used to avoid mosquito bites in Benin, were investigated. Methods Essential oils of nine plant species were extracted by hydrodistillation, and their chemical compositions were identified by GC-MS. These oils were tested on susceptible “kisumu” and resistant “ladji-Cotonou” strains of Anopheles gambiae, following WHO test procedures for insecticide resistance monitoring in malaria vector mosquitoes. Results Different chemical compositions were obtained from the essential oils of the plant species. The major constituents identified were as follows: neral and geranial for Cymbopogon citratus, Z-carveol, E-p-mentha-1(7),8-dien-2-ol and E-p-mentha-2,8-dienol for Cymbopogon giganteus, piperitone for Cymbopogon schoenanthus, citronellal and citronellol for Eucalyptus citriodora, p-cymene, caryophyllene oxide and spathulenol for Eucalyptus tereticornis, 3-tetradecanone for Cochlospermum tinctorium and Cochlospermum planchonii, methyl salicylate for Securidaca longepedunculata and ascaridole for Chenopodium ambrosioides. The diagnostic dose was 0.77% for C. citratus, 2.80% for E. tereticornis, 3.37% for E. citriodora, 4.26% for C. ambrosioides, 5.48% for C. schoenanthus and 7.36% for C. giganteus. The highest diagnostic doses were obtained with S. longepedunculata (9.84%), C. tinctorium (11.56%) and C. planchonii (15.22%), compared to permethrin 0.75%. A. gambiae cotonou, which is resistant to pyrethroids, showed significant tolerance to essential oils from C. tinctorium and S. longepedunculata as expected but was

Variant Ionotropic Receptors in the Malaria Vector Mosquito Anopheles gambiae Tuned to Amines and Carboxylic Acids

PubMed Central

Pitts, R. Jason; Derryberry, Stephen L.; Zhang, Zhiwei; Zwiebel, Laurence J.

2017-01-01

The principal Afrotropical human malaria vector mosquito, Anopheles gambiae, remains a significant threat to global health. A critical component in the transmission of malaria is the ability of An. gambiae females to detect and respond to human-derived chemical kairomones in their search for blood meal hosts. The basis for host odor responses resides in olfactory receptor neurons (ORNs) that express chemoreceptors encoded by large gene families, including the odorant receptors (ORs) and the variant ionotropic receptors (IRs). While ORs have been the focus of extensive investigation, functional IR complexes and the chemical compounds that activate them have not been identified in An. gambiae. Here we report the transcriptional profiles and functional characterization of three An. gambiae IR (AgIr) complexes that specifically respond to amines or carboxylic acids - two classes of semiochemicals that have been implicated in mediating host-seeking by adult females but are not known to activate An. gambiae ORs (AgOrs). Our results suggest that AgIrs play critical roles in the detection and behavioral responses to important classes of host odors that are underrepresented in the AgOr chemical space. PMID:28067294

A peroxidase/dual oxidase system modulates midgut epithelial immunity in Anopheles gambiae.

PubMed

Kumar, Sanjeev; Molina-Cruz, Alvaro; Gupta, Lalita; Rodrigues, Janneth; Barillas-Mury, Carolina

2010-03-26

Extracellular matrices in diverse biological systems are cross-linked by dityrosine covalent bonds catalyzed by the peroxidase/oxidase system. We show that a peroxidase, secreted by the Anopheles gambiae midgut, and dual oxidase form a dityrosine network that decreases gut permeability to immune elicitors. This network protects the microbiota by preventing activation of epithelial immunity. It also provides a suitable environment for malaria parasites to develop within the midgut lumen without inducing nitric oxide synthase expression. Disruption of this barrier results in strong and effective pathogen-specific immune responses.

The genetics of inviability and male sterility in hybrids between Anopheles gambiae and An. arabiensis.

PubMed

Slotman, M; Della Torre, A; Powell, J R

2004-05-01

Male hybrids between Anopheles gambiae and An. arabiensis suffer from hybrid sterility, and inviability effects are sometimes present as well. We examined the genetic basis of these reproductive barriers between the two species, using 21 microsatellite markers. Generally, recessive inviability effects were found on the X chromosome of gambiae that are incompatible with at least one factor on each arabiensis autosome. Inviability is complete when the gambiae and arabiensis inviability factors are hemi- or homozygous. Using a QTL mapping approach, regions that contribute to male hybrid sterility were also identified. The X chromosome has a disproportionately large effect on male hybrid sterility. Additionally, several moderate-to-large autosomal QTL were found in both species. The effect of these autosomal QTL is contingent upon the presence of an X chromosome from the other species. Substantial regions of the autosomes do not contribute markedly to male hybrid sterility. Finally, no evidence for epistatic interactions between conspecific sterility loci was found.

1.45 Å resolution structure of SRPN18 from the malaria vector Anopheles gambiae

PubMed Central

Meekins, David A.; Zhang, Xin; Battaile, Kevin P.; Lovell, Scott; Michel, Kristin

2016-01-01

Serine protease inhibitors (serpins) in insects function within development, wound healing and immunity. The genome of the African malaria vector, Anopheles gambiae, encodes 23 distinct serpin proteins, several of which are implicated in disease-relevant physiological responses. A. gambiae serpin 18 (SRPN18) was previously categorized as non-inhibitory based on the sequence of its reactive-center loop (RCL), a region responsible for targeting and initiating protease inhibition. The crystal structure of A. gambiae SRPN18 was determined to a resolution of 1.45 Å, including nearly the entire RCL in one of the two molecules in the asymmetric unit. The structure reveals that the SRPN18 RCL is extremely short and constricted, a feature associated with noncanonical inhibitors or non-inhibitory serpin superfamily members. Furthermore, the SRPN18 RCL does not contain a suitable protease target site and contains a large number of prolines. The SRPN18 structure therefore reveals a unique RCL architecture among the highly conserved serpin fold. PMID:27917832

Anopheles gambiae Purine Nucleoside Phosphorylase: Catalysis, Structure, and Inhibition

DOE Office of Scientific and Technical Information (OSTI.GOV)

Taylor,E.; Rinaldo-Matthis, A.; Li, L.

The purine salvage pathway of Anopheles gambiae, a mosquito that transmits malaria, has been identified in genome searches on the basis of sequence homology with characterized enzymes. Purine nucleoside phosphorylase (PNP) is a target for the development of therapeutic agents in humans and purine auxotrophs, including malarial parasites. The PNP from Anopheles gambiae (AgPNP) was expressed in Escherichia coli and compared to the PNPs from Homo sapiens (HsPNP) and Plasmodium falciparum (PfPNP). AgPNP has kcat values of 54 and 41 s-1 for 2'-deoxyinosine and inosine, its preferred substrates, and 1.0 s-1 for guanosine. However, the chemical step is fast formore » AgPNP at 226 s-1 for guanosine in pre-steady-state studies. 5'-Deaza-1'-aza-2'-deoxy-1'-(9-methylene)-Immucillin-H (DADMe-ImmH) is a transition-state mimic for a 2'-deoxyinosine ribocation with a fully dissociated N-ribosidic bond and is a slow-onset, tight-binding inhibitor with a dissociation constant of 3.5 pM. This is the tightest-binding inhibitor known for any PNP, with a remarkable Km/Ki* of 5.4 x 107, and is consistent with enzymatic transition state predictions of enhanced transition-state analogue binding in enzymes with enhanced catalytic efficiency. Deoxyguanosine is a weaker substrate than deoxyinosine, and DADMe-Immucillin-G is less tightly bound than DADMe-ImmH, with a dissociation constant of 23 pM for AgPNP as compared to 7 pM for HsPNP. The crystal structure of AgPNP was determined in complex with DADMe-ImmH and phosphate to a resolution of 2.2 Angstroms to reveal the differences in substrate and inhibitor specificity. The distance from the N1' cation to the phosphate O4 anion is shorter in the AgPNP{center_dot}DADMe-ImmH{center_dot}PO4 complex than in HsPNP{center_dot}DADMe-ImmH{center_dot}SO4, offering one explanation for the stronger inhibitory effect of DADMe-ImmH for AgPNP.« less

Effect of naturally occurring Wolbachia in Anopheles gambiae s.l. mosquitoes from Mali on Plasmodium falciparum malaria transmission

PubMed Central

Gomes, Fabio M.; Hixson, Bretta L.; Tyner, Miles D. W.; Ramirez, Jose Luis; Canepa, Gaspar E.; Alves e Silva, Thiago Luiz; Molina-Cruz, Alvaro; Keita, Moussa; Kane, Fouseyni; Traoré, Boïssé; Sogoba, Nafomon; Barillas-Mury, Carolina

2017-01-01

A naturally occurring Wolbachia strain (wAnga-Mali) was identified in mosquitoes of the Anopheles gambiae complex collected in the Malian villages of Dangassa and Kenieroba. Phylogenetic analysis of the nucleotide sequence of two 16S rRNA regions showed that wAnga-Mali clusters with Wolbachia strains from supergroup A and has the highest homology to a Wolbachia strain isolated from cat fleas (Ctenocephalides). wAnga-Mali is different from two Wolbachia strains previously reported in A. gambiae from Burkina Faso (wAnga_VK5_STP and wAnga_VK5_3.1a). Quantitative analysis of Wolbachia and Plasmodium sporozoite infection in field-collected mosquitoes indicates that the prevalence and intensity of Plasmodium falciparum sporozoite infection is significantly lower in Wolbachia-infected females. The presence of Wolbachia in females from a laboratory Anopheles coluzzii (A. gambiae, M form) colony experimentally infected with P. falciparum (NF54 strain) gametocyte cultures slightly enhanced oocyst infection. However, Wolbachia infection significantly reduced the prevalence and intensity of sporozoite infection, as observed in the field. This indicates that wAnga-Mali infection does not limit early stages of Plasmodium infection in the mosquito, but it has a strong deleterious effect on sporozoites and reduces malaria transmission. PMID:29114059

Development of vegetable farming: a cause of the emergence of insecticide resistance in populations of Anopheles gambiae in urban areas of Benin.

PubMed

Yadouleton, Anges William M; Asidi, Alex; Djouaka, Rousseau F; Braïma, James; Agossou, Christian D; Akogbeto, Martin C

2009-05-14

A fast development of urban agriculture has recently taken place in many areas in the Republic of Benin. This study aims to assess the rapid expansion of urban agriculture especially, its contribution to the emergence of insecticide resistance in populations of Anopheles gambiae. The protocol was based on the collection of sociological data by interviewing vegetable farmers regarding various agricultural practices and the types of pesticides used. Bioassay tests were performed to assess the susceptibility of malaria vectors to various agricultural insecticides and biochemical analysis were done to characterize molecular status of population of An. gambiae. This research showed that:(1) The rapid development of urban agriculture is related to unemployment observed in cities, rural exodus and the search for a balanced diet by urban populations;(2) Urban agriculture increases the farmers' household income and their living standard;(3) At a molecular level, PCR revealed the presence of three sub-species of An. gambiae (An. gambiae s.s., Anopheles melas and Anopheles arabiensis) and two molecular forms (M and S). The kdr west mutation recorded in samples from the three sites and more specifically on the M forms seems to be one of the major resistance mechanisms found in An. gambiae from agricultural areas. Insecticide susceptibility tests conducted during this research revealed a clear pattern of resistance to permethrin (76% mortality rate at Parakou; 23.5% at Porto-Novo and 17% at Cotonou). This study confirmed an increase activity of the vegetable farming in urban areas of Benin. This has led to the use of insecticide in an improper manner to control vegetable pests, thus exerting a huge selection pressure on mosquito larval population, which resulted to the emergence of insecticide resistance in malaria vectors.

Expression of metallothionein and α-tubulin in heavy metal-tolerant Anopheles gambiae sensu stricto (Diptera: Culicidae)

PubMed Central

Mireji, Paul O.; Keating, Joseph; Hassanali, Ahmed; Impoinvil, Daniel E.; Mbogo, Charles M.; Njeri, Martha; Nyambaka, Hudson; Kenya, Eucharia; Githure, John I; Beier, John C.

2009-01-01

Anopheles mosquitoes have been shown to adapt to heavy metals in their natural habitats. In this study we explored the possibility of using Anopheles gambiae sensu stricto as bio-reporters for environmental heavy metal pollution through expressions of their metal responsive metallothionein and α-tubulin genes. The study was undertaken with third instar larvae after selection by cadmium, copper, or lead at LC30 through five successive generations. Expression levels were determined in the fifth generation by semi quantitative RT-PCR on the experimental and control populations. The data were analyzed using one-way ANOVA. The highest metallothionein (F3, 11= 4.574, P = 0.038) and α-tubulin (F3,11= 12.961, P = 0.002) responses were observed in cadmium-tolerant treatments. There was significantly higher expression of metallothionein in cadmium or copper treatments relative to the control (P = 0.012), and in cadmium than in lead treatments (P = 0.044). Expressions of α-tubulin were significantly higher in cadmium than in control treatments (P = 0.008). These results demonstrate capacity of An. gambiae s.s. to develop tolerance to increased levels of heavy metal challenge. The results also confirm the potential of heavy metal responsive genes in mosquitoes as possible bio-indicators of heavy metal environmental pollution. How the tolerance and expressions relate to An. gambiae s.s. fitness and vectorial capacity in the environment remains to be elucidated. PMID:19735939

Visualizing Non Infectious and Infectious Anopheles gambiae Blood Feedings in Naive and Saliva-Immunized Mice

PubMed Central

Choumet, Valerie; Attout, Tarik; Chartier, Loïc; Khun, Huot; Sautereau, Jean; Robbe-Vincent, Annie; Brey, Paul; Huerre, Michel; Bain, Odile

2012-01-01

Background Anopheles gambiae is a major vector of malaria and lymphatic filariasis. The arthropod-host interactions occurring at the skin interface are complex and dynamic. We used a global approach to describe the interaction between the mosquito (infected or uninfected) and the skin of mammals during blood feeding. Methods Intravital video microscopy was used to characterize several features during blood feeding. The deposition and movement of Plasmodium berghei sporozoites in the dermis were also observed. We also used histological techniques to analyze the impact of infected and uninfected feedings on the skin cell response in naive mice. Results The mouthparts were highly mobile within the skin during the probing phase. Probing time increased with mosquito age, with possible effects on pathogen transmission. Repletion was achieved by capillary feeding. The presence of sporozoites in the salivary glands modified the behavior of the mosquitoes, with infected females tending to probe more than uninfected females (86% versus 44%). A white area around the tip of the proboscis was observed when the mosquitoes fed on blood from the vessels of mice immunized with saliva. Mosquito feedings elicited an acute inflammatory response in naive mice that peaked three hours after the bite. Polynuclear and mast cells were associated with saliva deposits. We describe the first visualization of saliva in the skin by immunohistochemistry (IHC) with antibodies directed against saliva. Both saliva deposits and sporozoites were detected in the skin for up to 18 h after the bite. Conclusion This study, in which we visualized the probing and engorgement phases of Anopheles gambiae blood meals, provides precise information about the behavior of the insect as a function of its infection status and the presence or absence of anti-saliva antibodies. It also provides insight into the possible consequences of the inflammatory reaction for blood feeding and pathogen transmission. PMID

Toxicity of six plant extracts and two pyridine alkaloids from Ricinus communis against the malaria vector Anopheles gambiae

USDA-ARS?s Scientific Manuscript database

The African malaria vector, Anopheles gambiae s.s., is known to feed selectively on certain plants for sugar sources. However, the adaptive significance of this behavior especially on how the extracts of such plants impact on the fitness of this vector has not been explored. This study determined th...

Insecticide resistance mechanisms associated with different environments in the malaria vector Anopheles gambiae: a case study in Tanzania

PubMed Central

2014-01-01

Background Resistance of mosquitoes to insecticides is a growing concern in Africa. Since only a few insecticides are used for public health and limited development of new molecules is expected in the next decade, maintaining the efficacy of control programmes mostly relies on resistance management strategies. Developing such strategies requires a deep understanding of factors influencing resistance together with characterizing the mechanisms involved. Among factors likely to influence insecticide resistance in mosquitoes, agriculture and urbanization have been implicated but rarely studied in detail. The present study aimed at comparing insecticide resistance levels and associated mechanisms across multiple Anopheles gambiae sensu lato populations from different environments. Methods Nine populations were sampled in three areas of Tanzania showing contrasting agriculture activity, urbanization and usage of insecticides for vector control. Insecticide resistance levels were measured in larvae and adults through bioassays with deltamethrin, DDT and bendiocarb. The distribution of An. gambiae sub-species and pyrethroid target-site mutations (kdr) were investigated using molecular assays. A microarray approach was used for identifying transcription level variations associated to different environments and insecticide resistance. Results Elevated resistance levels to deltamethrin and DDT were identified in agriculture and urban areas as compared to the susceptible strain Kisumu. A significant correlation was found between adult deltamethrin resistance and agriculture activity. The subspecies Anopheles arabiensis was predominant with only few An. gambiae sensu stricto identified in the urban area of Dar es Salaam. The L1014S kdr mutation was detected at elevated frequency in An gambiae s.s. in the urban area but remains sporadic in An. arabiensis specimens. Microarrays identified 416 transcripts differentially expressed in any area versus the susceptible reference

Rodent malaria-resistant strains of the mosquito, Anopheles gambiae, have slower population growth than -susceptible strains

PubMed Central

Voordouw, Maarten J; Anholt, Bradley R; Taylor, Pam J; Hurd, Hilary

2009-01-01

Background Trade-offs between anti-parasite defence mechanisms and other life history traits limit the evolution of host resistance to parasites and have important implications for understanding diseases such as malaria. Mosquitoes have not evolved complete resistance to malaria parasites and one hypothesis is that anti-malaria defence mechanisms are costly. Results We used matrix population models to compare the population growth rates among lines of Anopheles gambiae that had been selected for resistance or high susceptibility to the rodent malaria parasite, Plasmodium yoelii nigeriensis. The population growth rate of the resistant line was significantly lower than that of the highly susceptible and the unselected control lines, regardless of whether mosquitoes were infected with Plasmodium or not. The lower population growth of malaria-resistant mosquitoes was caused by reduced post blood-feeding survival of females and poor egg hatching. Conclusion With respect to eradicating malaria, the strategy of releasing Plasmodium-resistant Anopheles mosquitoes is unlikely to be successful if the costs of Plasmodium-resistance in the field are as great as the ones measured in this study. High densities of malaria-resistant mosquitoes would have to be maintained by continuous release from captive breeding facilities. PMID:19379508

Characterization of hemocytes from the mosquitoes Anopheles gambiae and Aedes aegypti.

PubMed

Castillo, J C; Robertson, A E; Strand, M R

2006-12-01

Hemocytes are an essential component of the mosquito immune system but current knowledge of the types of hemocytes mosquitoes produce, their relative abundance, and their functions is limited. Addressing these issues requires improved methods for collecting and maintaining mosquito hemocytes in vitro, and comparative data that address whether important vector species produce similar or different hemocyte types. Toward this end, we conducted a comparative study with Anopheles gambiae and Aedes aegypti. Collection method greatly affected the number of hemocytes and contaminants obtained from adult females of each species. Using a collection method called high injection/recovery, we concluded that hemolymph from An. gambiae and Ae. aegypti adult females contains three hemocyte types (granulocytes, oenocytoids and prohemocytes) that were distinguished from one another by a combination of morphological and functional markers. Significantly more hemocytes were recovered from An. gambiae females than Ae. aegypti. However, granulocytes were the most abundant cell type in both species while oenocytoids and prohemocytes comprised less than 10% of the total hemocyte population. The same hemocyte types were collected from larvae, pupae and adult males albeit the absolute number and proportion of each hemocyte type differed from adult females. The number of hemocytes recovered from sugar fed females declined with age but blood feeding transiently increased hemocyte abundance. Two antibodies tested as potential hemocyte markers (anti-PP06 and anti-Dox-A2) also exhibited alterations in staining patterns following immune challenge with the bacterium Escherichia coli.

Laboratory evaluation of the irritancy of bendiocarb, lambda-cyhalothrin and DDT to Anopheles gambiae.

PubMed

Evans, R G

1993-09-01

In a laboratory study, the irritancy of bendiocarb, lambda-cyhalothrin and DDT to Anopheles gambiae was evaluated at field, 1/3 field and 1/10 field rates using WHO conical exposure chambers and excito-repellency test boxes. Bendiocarb was the least irritant insecticide at all rates, inducing levels of takeoff, flight and exiting behavior similar to those of a distilled water control treatment. Of those mosquitoes introduced to the bendiocarb-treated boxes, not more than 1% exited and survived at any dose rate. Lambda-cyhalothrin and DDT were highly irritant to An. gambiae, inducing a strong stimulation to take off and fly and also a high level of exiting. Exiting-survival rates associated with lambda-cyhalothrin and DDT were between 15 and 51%. The relevance of these findings to the control of mosquito populations and the prevention of malaria transmission is discussed.

Deforestation and Vectorial Capacity of Anopheles gambiae Giles Mosquitoes in Malaria Transmission, Kenya

PubMed Central

Afrane, Yaw A.; Little, Tom J.; Lawson, Bernard W.; Githeko, Andrew K.

2008-01-01

We investigated the effects of deforestation on microclimates and sporogonic development of Plasmodium falciparum parasites in Anopheles gambiae mosquitoes in an area of the western Kenyan highland prone to malaria epidemics. An. gambiae mosquitoes were fed with P. falciparum–infected blood through membrane feeders. Fed mosquitoes were placed in houses in forested and deforested areas in a highland area (1,500 m above sea level) and monitored for parasite development. Deforested sites had higher temperatures and relative humidities, and the overall infection rate of mosquitoes was increased compared with that in forested sites. Sporozoites appeared on average 1.1 days earlier in deforested areas. Vectorial capacity was estimated to be 77.7% higher in the deforested site than in the forested site. We showed that deforestation changes microclimates, leading to more rapid sporogonic development of P. falciparum and to a marked increase of malaria risk in the western Kenyan highland. PMID:18826815

Chloroquine mediated modulation of Anopheles gambiae gene expression.

PubMed

Abrantes, Patrícia; Dimopoulos, George; Grosso, Ana Rita; do Rosário, Virgílio E; Silveira, Henrique

2008-07-02

Plasmodium development in the mosquito is crucial for malaria transmission and depends on the parasite's interaction with a variety of cell types and specific mosquito factors that have both positive and negative effects on infection. Whereas the defensive response of the mosquito contributes to a decrease in parasite numbers during these stages, some components of the blood meal are known to favor infection, potentiating the risk of increased transmission. The presence of the antimalarial drug chloroquine in the mosquito's blood meal has been associated with an increase in Plasmodium infectivity for the mosquito, which is possibly caused by chloroquine interfering with the capacity of the mosquito to defend against the infection. In this study, we report a detailed survey of the Anopheles gambiae genes that are differentially regulated by the presence of chloroquine in the blood meal, using an A. gambiae cDNA microarray. The effect of chloroquine on transcript abundance was evaluated separately for non-infected and Plasmodium berghei-infected mosquitoes. Chloroquine was found to affect the abundance of transcripts that encode proteins involved in a variety of processes, including immunity, apoptosis, cytoskeleton and the response to oxidative stress. This pattern of differential gene expression may explain the weakened mosquito defense response which accounts for the increased infectivity observed in chloroquine-treated mosquitoes. The results of the present study suggest that chloroquine can interfere with several putative mosquito mechanisms of defense against Plasmodium at the level of gene expression and highlight the need for a better understanding of the impacts of antimalarial agents on parasite transmission.

Insecticide resistance status of three malaria vectors, Anopheles gambiae (s.l.), An. funestus and An. mascarensis, from the south, central and east coasts of Madagascar.

PubMed

Rakotoson, Jean-Desire; Fornadel, Christen M; Belemvire, Allison; Norris, Laura C; George, Kristen; Caranci, Angela; Lucas, Bradford; Dengela, Dereje

2017-08-23

Insecticide-based vector control, which comprises use of insecticide-treated bed nets (ITNs) and indoor residual spraying (IRS), is the key method to malaria control in Madagascar. However, its effectiveness is threatened as vectors become resistant to insecticides. This study investigated the resistance status of malaria vectors in Madagascar to various insecticides recommended for use in ITNs and/or IRS. WHO tube and CDC bottle bioassays were performed on populations of Anopheles gambiae (s.l.), An. funestus and An. mascarensis. Adult female An. gambiae (s.l.) mosquitoes reared from field-collected larvae and pupae were tested for their resistance to DDT, permethrin, deltamethrin, alpha-cypermethrin, lambda-cyhalothrin, bendiocarb and pirimiphos-methyl. Resting An. funestus and An. mascarensis female mosquitoes collected from unsprayed surfaces were tested against permethrin, deltamethrin and pirimiphos-methyl. The effect on insecticide resistance of pre-exposure to the synergists piperonyl-butoxide (PBO) and S,S,S-tributyl phosphorotrithioate (DEF) also was assessed. Molecular analyses were done to identify species and determine the presence of knock-down resistance (kdr) and acetylcholinesterase resistance (ace-1 R ) gene mutations. Anopheles funestus and An. mascarensis were fully susceptible to permethrin, deltamethrin and pirimiphos-methyl. Anopheles gambiae (s.l.) was fully susceptible to bendiocarb and pirimiphos-methyl. Among the 17 An. gambiae (s.l.) populations tested for deltamethrin, no confirmed resistance was recorded, but suspected resistance was observed in two sites. Anopheles gambiae (s.l.) was resistant to permethrin in four out of 18 sites (mortality 68-89%) and to alpha-cypermethrin (89% mortality) and lambda-cyhalothrin (80% and 85%) in one of 17 sites, using one or both assay methods. Pre-exposure to PBO restored full susceptibility to all pyrethroids tested except in one site where only partial restoration to permethrin was observed. DEF

Modelling the impact of vector control interventions on Anopheles gambiae population dynamics

PubMed Central

2011-01-01

Background Intensive anti-malaria campaigns targeting the Anopheles population have demonstrated substantial reductions in adult mosquito density. Understanding the population dynamics of Anopheles mosquitoes throughout their whole lifecycle is important to assess the likely impact of vector control interventions alone and in combination as well as to aid the design of novel interventions. Methods An ecological model of Anopheles gambiae sensu lato populations incorporating a rainfall-dependent carrying capacity and density-dependent regulation of mosquito larvae in breeding sites is developed. The model is fitted to adult mosquito catch and rainfall data from 8 villages in the Garki District of Nigeria (the 'Garki Project') using Bayesian Markov Chain Monte Carlo methods and prior estimates of parameters derived from the literature. The model is used to compare the impact of vector control interventions directed against adult mosquito stages - long-lasting insecticide treated nets (LLIN), indoor residual spraying (IRS) - and directed against aquatic mosquito stages, alone and in combination on adult mosquito density. Results A model in which density-dependent regulation occurs in the larval stages via a linear association between larval density and larval death rates provided a good fit to seasonal adult mosquito catches. The effective mosquito reproduction number in the presence of density-dependent regulation is dependent on seasonal rainfall patterns and peaks at the start of the rainy season. In addition to killing adult mosquitoes during the extrinsic incubation period, LLINs and IRS also result in less eggs being oviposited in breeding sites leading to further reductions in adult mosquito density. Combining interventions such as the application of larvicidal or pupacidal agents that target the aquatic stages of the mosquito lifecycle with LLINs or IRS can lead to substantial reductions in adult mosquito density. Conclusions Density-dependent regulation of

Effects of Plant-Community Composition on the Vectorial Capacity and Fitness of the Malaria Mosquito Anopheles gambiae

PubMed Central

Stone, Christopher M.; Jackson, Bryan T.; Foster, Woodbridge A.

2012-01-01

Dynamics of Anopheles gambiae abundance and malaria transmission potential rely strongly on environmental conditions. Female and male An. gambiae use sugar and are affected by its absence, but how the presence or absence of nectariferous plants affects An. gambiae abundance and vectorial capacity has not been studied. We report on four replicates of a cohort study performed in mesocosms with sugar-poor and sugar-rich plants, in which we measured mosquito survival, biting rates, and fecundity. Survivorship was greater with access to sugar-rich plant species, and mortality patterns were age-dependent. Sugar-poor populations experienced Weibull mortality patterns, and of four populations in the sugar-rich environment, two female and three male subpopulations were better fitted by Gompertz functions. A tendency toward higher biting rates in sugar-poor mesocosms, particularly for young females, was found. Therefore, vectorial capacity was pulled in opposing directions by nectar availability, resulting in highly variable vectorial capacity values. PMID:22927493

Life on the edge: African malaria mosquito (Anopheles gambiae s. l.) larvae are amphibious.

PubMed

Miller, James R; Huang, Juan; Vulule, John; Walker, Edward D

2007-03-01

Anopheles gambiae s.l. is the main vector of malaria in Sub-Saharan Africa. Here, an estimated 1 million people die every year from this disease. Despite considerable research on An. gambiae that increasingly explores sub-organismal phenomena, important facets of the field biology of this deadly insect are yet being discovered. In the current study, we used simple observational tools to reveal that the habitat of larval An. gambiae is not limited within the boundaries of temporary mud puddles, as has been the accepted generalization. Thus, control tactics aimed at immatures must consider zones larger than puddles per se. In fact, eggs are more likely to be found outside than inside puddles. Eggs can develop and larvae can emerge on mud. Larvae are then capable of three distinct modes of terrestrial displacement (two active and one passive), whereby, they can reach standing water. On mud bearing a film of water, larvae actively displace backwards by sinusoidal undulations shown to be only a slight variation of the swimming motor program. On drying mud, larvae switch to a slower and forward form of active locomotion resembling that of a crawling caterpillar. During rains, small larvae may be passively displaced by flowing rainwater so as to be deposited into puddles. These capabilities for being amphibious, along with very rapid growth and development, help explain how An. gambiae thrives in a highly uncertain and often hostile larval environment.

Molecular evolution of the three short PGRPs of the malaria vectors Anopheles gambiae and Anopheles arabiensis in East Africa.

PubMed

Mendes, Cristina; Felix, Rute; Sousa, Ana-Margarida; Lamego, Joana; Charlwood, Derek; do Rosário, Virgílio E; Pinto, João; Silveira, Henrique

2010-01-12

Immune responses to parasites, which start with pathogen recognition, play a decisive role in the control of the infection in mosquitoes. Peptidoglycan recognition proteins (PGRPs) are an important family of pattern recognition receptors that are involved in the activation of these immune reactions. Pathogen pressure can exert adaptive changes in host genes that are crucial components of the vector's defence. The aim of this study was to determine the molecular evolution of the three short PGRPs (PGRP-S1, PGRP-S2 and PGRP-S3) in the two main African malaria vectors - Anopheles gambiae and Anopheles arabiensis. Genetic diversity of An. gambiae and An. arabiensis PGRP-S1, PGRP-S2 and PGRP-S3 was investigated in samples collected from Mozambique and Tanzania. PGRP-S1 diversity was lower than for PGRP-S2 and PGRP-S3. PGRP-S1 was the only gene differentiated between the two species. All the comparisons made for PGRP-S1 showed significant P-values for Fst estimates and AMOVA confirming a clear separation between species. For PGRP-S2 and PGRP-S3 genes it was not possible to group populations either by species or by geographic region. Phylogenetic networks reinforced the results obtained by the AMOVA and Fst values. The ratio of nonsynonymous substitutions (Ka)/synonymous substitutions (Ks) for the duplicate pair PGRP-S2 and PGRP-S3 was very similar and lower than 1. The 3D model of the different proteins coded by these genes showed that amino acid substitutions were concentrated at the periphery of the protein rather than at the peptidoglycan recognition site. PGRP-S1 is less diverse and showed higher divergence between An. gambiae and An. arabiensis regardless of geographic location. This probably relates to its location in the chromosome-X, while PGRP-S2 and PGRP-S3, located in chromosome-2L, showed signs of autosomal introgression. The two short PGRP genes located in the chromosome-2L were under purifying selection, which suggests functional constraints. Different

Evaluating the effect of postmating isolation between molecular forms of Anopheles gambiae (Diptera: Culicidae).

PubMed

Diabaté, Abdoulaye; Dabire, Roch K; Millogo, Niama; Lehmann, Tovi

2007-01-01

Multiple families representing all possible combinations of crosses between the two molecular forms of Anopheles gambiae sensu stricto Giles and their hybrids were set up using forced mating between offspring of wild-collected females. The results showed that the reproductive output of hybrids and their backcrosses was similar to that of the pure forms as measured by egg batch size, hatching rate, and larval development success. No sex ratio distortion was found among the offspring. We concluded that postmating developmental barriers do not contribute to the isolation between the molecular forms.

Mosaic: a position-effect variegation eye-color mutant in the mosquito Anopheles gambiae.

PubMed

Benedict, M Q; McNitt, L M; Cornel, A J; Collins, F H

2000-01-01

The Mosaic (Mos) mutation, isolated in the F1 of 60Co-irradiated mosquitoes, confers variegated eye color to third and fourth instar larvae, pupae, and adults of the mosquito Anopheles gambiae. Mos is recessive in wild pink eye (p+) individuals, but is dominant and confers areas of wild-type pigment in mutant pink eye backgrounds. Mos is located 14.4 cM from pink eye on the X chromosome and is associated with a duplication of division 2B euchromatin that has been inserted into division 6 heterochromatin. Various combinations of Mos, pink eye alleles, and the autosomal mutation red eye were produced. In all cases, the darker pigmented regions of the eye in Mos individuals show the phenotypic interactions expected if the phenotype of those regions is due to expression of a p+ allele. Expression of Mos is suppressed by rearing larvae at 32 degrees C relative to 22 degrees C. All of these characteristics are consistent with Mos being a duplicated wild copy of the pink eye gene undergoing position-effect variegation.

1.45 Å resolution structure of SRPN18 from the malaria vector Anopheles gambiae

DOE Office of Scientific and Technical Information (OSTI.GOV)

Meekins, David A.; Zhang, Xin; Battaile, Kevin P.

Serine protease inhibitors (serpins) in insects function within development, wound healing and immunity. The genome of the African malaria vector,Anopheles gambiae, encodes 23 distinct serpin proteins, several of which are implicated in disease-relevant physiological responses.A. gambiaeserpin 18 (SRPN18) was previously categorized as non-inhibitory based on the sequence of its reactive-center loop (RCL), a region responsible for targeting and initiating protease inhibition. The crystal structure ofA. gambiaeSRPN18 was determined to a resolution of 1.45 Å, including nearly the entire RCL in one of the two molecules in the asymmetric unit. The structure reveals that the SRPN18 RCL is extremely short andmore » constricted, a feature associated with noncanonical inhibitors or non-inhibitory serpin superfamily members. Furthermore, the SRPN18 RCL does not contain a suitable protease target site and contains a large number of prolines. The SRPN18 structure therefore reveals a unique RCL architecture among the highly conserved serpin fold.« less

'Repel all biters': an enhanced collection of endophilic Anopheles gambiae and Anopheles arabiensis in CDC light-traps, from the Kagera Region of Tanzania, in the presence of a combination mosquito net impregnated with piperonyl butoxide and permethrin.

PubMed

LeClair, Corey; Cronery, Judith; Kessy, Enock; Tomás, Elsa V E; Kulwa, Yohannes; Mosha, Franklin W; Rowland, Mark; Protopopoff, Natacha; Derek Charlwood, J

2017-08-15

Mosquito nets containing synergists designed to overcome metabolic resistance mechanisms in vectors have been developed. These may enhance excitability in the mosquitoes and affect how they respond to CDC light-traps. Investigating the behaviour of vectors of disease in relation to novel mosquito nets is, therefore, essential for the design of sampling and surveillance systems. In an initial experiment in Muleba, Tanzania, nine bedrooms from three housing clusters were sampled. CDC light-traps were operated indoors next to occupied untreated nets (UTN), Olyset ® long lasting insecticidal net (LLIN) and Olyset Plus ® LLIN containing piperonyl butoxide (PBO) synergist. Nets were rotated daily between the nine rooms over nine nights. A further series of experiments using the nets on alternate nights in a single room was undertaken during the short rains. Anopheles gambiae s.l. were collected in CDC light-traps, a window-trap and Furvela tent-trap. Anopheles gambiae s.l. were identified to species by polymerase chain reaction (PCR). In the initial experiment 97.7% of the 310 An. gambiae s.l. were An. gambiae s.s., the remainder being Anopheles arabiensis. The number of mosquitoes collected from 81 light-trap collections was greater in the presence of an Olyset [density rate ratio 1.81, 95% CI (1.22-2.67), p = 0.003] relative to an UTN. In a second experiment, in the wet season 84% of the 180 An. gambiae s.l. identified were An. arabiensis. The number of An. gambiae s.l. collected from a light-trap compared to a tent-trap was significantly higher when an Olyset Plus net was used compared to an UTN. Survival of the mosquitoes in the window trap was not reduced by the use of an Olyset Plus net in the bedroom relative to an Olyset net. Mosquitoes entering bedrooms, even those susceptible to pyrethroids, were not killed by contact with an Olyset Plus LLIN. The enhanced numbers of An. gambiae or An. arabiensis collected in light-traps when a treated net is used requires

A retrotransposable element from the mosquito Anopheles gambiae .

PubMed Central

Besansky, N J

1990-01-01

A family of middle repetitive elements from the African malaria vector Anopheles gambiae is described. Approximately 100 copies of the element, designated T1Ag, are dispersed in the genome. Full-length elements are 4.6 kilobase pairs in length, but truncation of the 5' end is common. Nucleotide sequences of one full-length, two 5'-truncated, and two 5' ends of T1Ag elements were determined and aligned to define a consensus sequence. Sequence analysis revealed two long, overlapping open reading frames followed by a polyadenylation signal, AATAAA, and a tail consisting of tandem repetitions of the motif TGAAA. No direct or inverted long terminal repeats (LTRs) were detected. The first open reading frame, 442 amino acids in length, includes a domain resembling that of nucleic acid-binding proteins. The second open reading frame, 975 amino acids long, resembles the reverse transcriptases of a category of retrotransposable elements without LTRs, variously termed class II retrotransposons, class III elements or non-LTR retrotransposons. Similarity at the sequence and structural levels places T1Ag in this category. Images PMID:1689457

Assessment of Anopheles salivary antigens as individual exposure biomarkers to species-specific malaria vector bites

PubMed Central

2012-01-01

Background Malaria transmission occurs during the blood feeding of infected anopheline mosquitoes concomitant with a saliva injection into the vertebrate host. In sub-Saharan Africa, most malaria transmission is due to Anopheles funestus s.s and to Anopheles gambiae s.l. (mainly Anopheles gambiae s.s. and Anopheles arabiensis). Several studies have demonstrated that the immune response against salivary antigens could be used to evaluate individual exposure to mosquito bites. The aim of this study was to assess the use of secreted salivary proteins as specific biomarkers of exposure to An. gambiae and/or An. funestus bites. Methods For this purpose, salivary gland proteins 6 (SG6) and 5′nucleotidases (5′nuc) from An. gambiae (gSG6 and g-5′nuc) and An. funestus (fSG6 and f-5′nuc) were selected and produced in recombinant form. The specificity of the IgG response against these salivary proteins was tested using an ELISA with sera from individuals living in three Senegalese villages (NDiop, n = 50; Dielmo, n = 38; and Diama, n = 46) that had been exposed to distinct densities and proportions of the Anopheles species. Individuals who had not been exposed to these tropical mosquitoes were used as controls (Marseille, n = 45). Results The IgG responses against SG6 recombinant proteins from these two Anopheles species and against g-5′nucleotidase from An. gambiae, were significantly higher in Senegalese individuals compared with controls who were not exposed to specific Anopheles species. Conversely, an association was observed between the level of An. funestus exposure and the serological immune response levels against the f-5′nucleotidase protein. Conclusion This study revealed an Anopheles salivary antigenic protein that could be considered to be a promising antigenic marker to distinguish malaria vector exposure at the species level. The epidemiological interest of such species-specific antigenic markers is discussed. PMID:23276246

Characterization of the c-type lysozyme gene family in Anopheles gambiae.

PubMed

Li, Bin; Calvo, Eric; Marinotti, Osvaldo; James, Anthony A; Paskewitz, Susan M

2005-11-07

Seven new c-type lysozyme genes were found using the Anopheles gambiae genome sequence, increasing to eight the total number of genes in this family identified in this species. The eight lysozymes in An. gambiae have considerable variation in gene structure and expression patterns. Lys c-6 has the most unusual primary amino acid structure as the predicted protein consists of five lysozyme-like domains. Transcript abundance of each c-type lysozyme was determined by semiquantitative RT-PCR. Lys c-1, c-6 and c-7 are expressed constitutively in all developmental stages from egg to adult. Lys c-2 and c-4 also are found in all stages, but with relatively much higher levels in adults. Conversely, Lys c-3 and c-8 transcripts are highest in larvae. Lys c-1, c-6 and c-7 transcripts are found in nearly all the adult tissue samples examined while Lys c-2 and Lys c-4 are more restricted in their expression. Lys c-1 and c-2 transcripts are clearly immune responsive and are increased significantly 6-12 h post challenge with bacteria. The functional adaptive changes that may have evolved during the expansion of this gene family are briefly discussed in terms of the expression patterns, gene and protein structures.

Status of pyrethroid resistance in Anopheles gambiae sensu lato.

PubMed Central

Chandre, F.; Darrier, F.; Manga, L.; Akogbeto, M.; Faye, O.; Mouchet, J.; Guillet, P.

1999-01-01

The present study confirms the presence of pyrethroid resistance among Anopheles gambiae s.l mosquitos in Côte d'Ivoire and reports the observation of such resistance in two other countries in West Africa (Benin and Burkina Faso). Malaria vector populations from Cameroon (Central Africa), Senegal (West Africa) and Botswana (southern Africa) were found to be susceptible to pyrethroids. In the most resistant mosquito populations, resistance to permethrin was associated with reduced mortality, not only with respect to this compound but also towards deltamethrin. Moreover, a significant increase in knockdown time was observed in some mosquito populations before any decrease in mortality, suggesting that knockdown time could be a good indicator for the early detection of pyrethroid resistance. In view of the current extension of such resistance, there is an urgent need to set up a network in Africa to evaluate its development. It is also vital that the impact of this resistance on pyrethroid-impregnated bednets be assessed. PMID:10212513

Laboratory tests of oviposition by the African malaria mosquito, Anopheles gambiae, on dark soil as influenced by presence or absence of vegetation

PubMed Central

Huang, Juan; Walker, Edward D; Otienoburu, Philip E; Amimo, Fred; Vulule, John; Miller, James R

2006-01-01

Background Physical objects like vegetation can influence oviposition by mosquitoes on soil or water substrates. Anopheles gambiae s. l. is generally thought to utilize puddles over bare soil as its prime larval habitat and to avoid standing water populated with vegetation. In Kisian, Kenya near Kisumu, water often pools in grassy drainage areas both during and after periods of infrequent rains, when typical puddle habitats become scarce because of drying. This raised the question of whether An. gambiae has the behavioural flexibility to switch ovipositional sites when puddles over bare soil are unavailable. Methods To test whether presence and height of grasses influenced oviposition, wild-caught gravid An. gambiae s. l. were offered paired choices between wet, bare soil and wet soil populated with mixed grasses or grasses of differing height. No-choice tests were also conducted by giving females either grassy soil or bare soil. Results In choice tests, females laid four times more eggs on bare, wet soil than soil populated with grasses. However in no-choice tests, egg output was not significantly different whether grasses were present or not. Females laid significantly more eggs on soil populated with short grass than with medium, or tall grass. Conclusion This work shows An. gambiae s. l. has the capacity to oviposit into grassy aquatic habitats when typical puddles over bare soil are unavailable. This knowledge will need to be considered in the design and implementation of programmes aimed at reducing malaria transmission by suppression of An. gambiae s. l. immatures. PMID:17038187

Extensive circadian and light regulation of the transcriptome in the malaria mosquito Anopheles gambiae

PubMed Central

2013-01-01

Background Mosquitoes exhibit 24 hr rhythms in flight activity, feeding, reproduction and development. To better understand the molecular basis for these rhythms in the nocturnal malaria vector Anopheles gambiae, we have utilized microarray analysis on time-of-day specific collections of mosquitoes over 48 hr to explore the coregulation of gene expression rhythms by the circadian clock and light, and compare these with the 24 hr rhythmic gene expression in the diurnal Aedes aegypti dengue vector mosquito. Results In time courses from An. gambiae head and body collected under light:dark cycle (LD) and constant dark (DD) conditions, we applied three algorithms that detect sinusoidal patterns and an algorithm that detects spikes in expression. This revealed across four experimental conditions 393 probes newly scored as rhythmic. These genes correspond to functions such as metabolic detoxification, immunity and nutrient sensing. This includes glutathione S-transferase GSTE5, whose expression pattern and chromosomal location are shared with other genes, suggesting shared chromosomal regulation; and pulsatile expression of the gene encoding CYP6M2, a cytochrome P450 that metabolizes pyrethroid insecticides. We explored the interaction of light and the circadian clock and highlight the regulation of odorant binding proteins (OBPs), important components of the olfactory system. We reveal that OBPs have unique expression patterns as mosquitoes make the transition from LD to DD conditions. We compared rhythmic expression between An. gambiae and Ae. aegypti heads collected under LD conditions using a single cosine fitting algorithm, and report distinct similarities and differences in the temporal regulation of genes involved in tRNA priming, the vesicular-type ATPase, olfaction and vision between the two species. Conclusions These data build on our previous analyses of time-of-day specific regulation of the An. gambiae transcriptome to reveal additional rhythmic genes, an

Influence of biological and physicochemical characteristics of larval habitats on the body size of Anopheles gambiae mosquitoes (Diptera: Culicidae) along the Kenyan coast

PubMed Central

Mwangangi, Joseph M.; Mbogo, Charles M.; Muturi, Ephantus J.; Nzovu, Joseph G.; Kabiru, Ephantus W.; Githure, John I.; Novak, Robert J.; Beier, John C.

2009-01-01

Background & objectives The number and productivity of larval habitats ultimately determine the density of adult mosquitoes. The biological and physicochemical conditions at the larval habitat affect larval development hence affecting the adult body size. The influence of biological and physicochemical characteristics on the body size of Anopheles gambiae was assessed in Jaribuni village, Kilifi district along the Kenyan Coast. Methods Ten cages measuring 1 × 1 × 1 m (1 m3) with a netting material were placed in 10 different aquatic habitats, which were positive for anopheline mosquito larvae. Emergent mosquitoes were collected daily by aspiration and the wing lengths were determined by microscopy. In the habitats, physicochemical parameters were assessed: pH, surface debris, algae and emergent plants, turbidity, substrate, nitrate, ammonia, phosphate and chlorophyll a content. Results A total of 685 anopheline and culicine mosquitoes were collected from the emergent cages. Only female mosquitoes were considered in this study. Among the Anopheles spp, 202 were An. gambiae s.s., eight An. arabiensis, two An. funestus, whereas the Culex spp was composed of 214 Cx. quinquefasciatus, 10 Cx. tigripes, eight Cx. annulioris and one Cx. cumminsii. The mean wing length of the female An. gambiae s.s. mosquitoes was 3.02 mm (n = 157), while that of An. arabiensis was 3.09 mm (n = 9). There were no associations between the wing lengths and the environmental and chemical parameters, except for a positive correlation between wing length of An. gambiae and chlorophyll a content (r = 0.622). The day on which the mosquitoes emerged was not significant for the anopheline (p = 0.324) or culicine mosquitoes (p = 0.374), because the mosquito emerged from the cages on a daily basis. Interpretation & conclusion In conclusion, there was variability in production of emergent mosquitoes from different habitats, which means that there should be targeted control on these habitats based on

3D Tracking of Mating Events in Wild Swarms of the Malaria Mosquito Anopheles gambiae

PubMed Central

Butail, Sachit; Manoukis, Nicholas; Diallo, Moussa; Yaro, Alpha S.; Dao, Adama; Traoré, Sekou F.; Ribeiro, José M.; Lehmann, Tovi; Paley, Derek A.

2013-01-01

We describe an automated tracking system that allows us to reconstruct the 3D kinematics of individual mosquitoes in swarms of Anopheles gambiae. The inputs to the tracking system are video streams recorded from a stereo camera system. The tracker uses a two-pass procedure to automatically localize and track mosquitoes within the swarm. A human-in-the-loop step verifies the estimates and connects broken tracks. The tracker performance is illustrated using footage of mating events filmed in Mali in August 2010. PMID:22254411

Ecological niche partitioning between Anopheles gambiae molecular forms in Cameroon: the ecological side of speciation

PubMed Central

Simard, Frédéric; Ayala, Diego; Kamdem, Guy Colince; Pombi, Marco; Etouna, Joachim; Ose, Kenji; Fotsing, Jean-Marie; Fontenille, Didier; Besansky, Nora J; Costantini, Carlo

2009-01-01

Background Speciation among members of the Anopheles gambiae complex is thought to be promoted by disruptive selection and ecological divergence acting on sets of adaptation genes protected from recombination by polymorphic paracentric chromosomal inversions. However, shared chromosomal polymorphisms between the M and S molecular forms of An. gambiae and insufficient information about their relationship with ecological divergence challenge this view. We used Geographic Information Systems, Ecological Niche Factor Analysis, and Bayesian multilocus genetic clustering to explore the nature and extent of ecological and chromosomal differentiation of M and S across all the biogeographic domains of Cameroon in Central Africa, in order to understand the role of chromosomal arrangements in ecological specialisation within and among molecular forms. Results Species distribution modelling with presence-only data revealed differences in the ecological niche of both molecular forms and the sibling species, An. arabiensis. The fundamental environmental envelope of the two molecular forms, however, overlapped to a large extent in the rainforest, where they occurred in sympatry. The S form had the greatest niche breadth of all three taxa, whereas An. arabiensis and the M form had the smallest niche overlap. Correspondence analysis of M and S karyotypes confirmed that molecular forms shared similar combinations of chromosomal inversion arrangements in response to the eco-climatic gradient defining the main biogeographic domains occurring across Cameroon. Savanna karyotypes of M and S, however, segregated along the smaller-scale environmental gradient defined by the second ordination axis. Population structure analysis identified three chromosomal clusters, each containing a mixture of M and S specimens. In both M and S, alternative karyotypes were segregating in contrasted environments, in agreement with a strong ecological adaptive value of chromosomal inversions. Conclusion Our

Daily oviposition patterns of the African malaria mosquito Anopheles gambiae Giles (Diptera: Culicidae) on different types of aqueous substrates

PubMed Central

Sumba, Leunita A; Okoth, Kenneth; Deng, Arop L; Githure, John; Knols, Bart GJ; Beier, John C; Hassanali, Ahmed

2004-01-01

Background Anopheles gambiae Giles is the most important vector of human malaria in sub-Saharan Africa. Knowledge of the factors that influence its daily oviposition pattern is crucial if field interventions targeting gravid females are to be successful. This laboratory study investigated the effect of oviposition substrate and time of blood feeding on daily oviposition patterns of An. gambiae mosquitoes. Methods Greenhouse-reared gravid and hypergravid (delayed oviposition onset) An. gambiae sensu stricto and wild-caught An. gambiae sensu lato were exposed to three types of substrates in choice and no-choice cage bioassays: water from a predominantly anopheline colonised ground pool (anopheline habitat water), swamp water mainly colonised by culicine larvae (culicine habitat water) and distilled water. The daily oviposition pattern and the number of eggs oviposited on each substrate during the entire egg-laying period were determined. The results were subjected to analysis of variance using the General Linear Model (GLM) procedure. Results The main oviposition time for greenhouse-reared An. gambiae s.s. was between 19:00 and 20:00 hrs, approximately one hour after sunset. Wild-caught gravid An. gambiae s.l. displayed two distinct peak oviposition times between 19:00 and 20:00 hrs and between 22:00 and 23:00 hrs, respectively. During these times, both greenhouse-reared and wild-caught mosquitoes significantly (P < 0.05) preferred anopheline habitat water to the culicine one. Peak oviposition activity was not delayed when the mosquitoes were exposed to the less preferred oviposition substrate (culicine habitat water). However, culicine water influenced negatively (P < 0.05) not only the number of eggs oviposited by the mosquitoes during peak oviposition time but also the overall number of gravid mosquitoes that laid their eggs on it. The differences in mosquito feeding times did not affect the daily oviposition patterns displayed. Conclusion This study shows that

Inversion 2La is associated with enhanced desiccation resistance in Anopheles gambiae

PubMed Central

Gray, Emilie M; Rocca, Kyle AC; Costantini, Carlo; Besansky, Nora J

2009-01-01

Background Anopheles gambiae, the principal vector of malignant malaria in Africa, occupies a wide range of habitats. Environmental flexibility may be conferred by a number of chromosomal inversions non-randomly associated with aridity, including 2La. The purpose of this study was to determine the physiological mechanisms associated with the 2La inversion that may result in the preferential survival of its carriers in hygrically-stressful environments. Methods Two homokaryotypic populations of A. gambiae (inverted 2La and standard 2L+a) were created from a parental laboratory colony polymorphic for 2La and standard for all other known inversions. Desiccation resistance, water, energy and dry mass of adult females of both populations were compared at several ages and following acclimation to a more arid environment. Results Females carrying 2La were significantly more resistant to desiccation than 2L+a females at emergence and four days post-emergence, for different reasons. Teneral 2La females had lower rates of water loss than their 2L+a counterparts, while at four days, 2La females had higher initial water content. No differences in desiccation resistance were found at eight days, with or without acclimation. However, acclimation resulted in both populations significantly reducing their rates of water loss and increasing their desiccation resistance. Acclimation had contrasting effects on the body characteristics of the two populations: 2La females boosted their glycogen stores and decreased lipids, whereas 2La females did the contrary. Conclusion Variation in rates of water loss and response to acclimation are associated with alternative arrangements of the 2La inversion. Understanding the mechanisms underlying these traits will help explain how inversion polymorphisms permit exploitation of a heterogeneous environment by this disease vector. PMID:19772577

Characterisation of Anopheles gambiae heme oxygenase and metalloporphyrin feeding suggests a potential role in reproduction.

PubMed

Spencer, Christopher S; Yunta, Cristina; de Lima, Glauber Pacelli Gomes; Hemmings, Kay; Lian, Lu-Yun; Lycett, Gareth; Paine, Mark J I

2018-05-03

The mosquito Anopheles gambiae is the principal vector for malaria in sub-Saharan Africa. The ability of A. gambiae to transmit malaria is strictly related to blood feeding and digestion, which releases nutrients for oogenesis, as well as substantial amounts of highly toxic free heme. Heme degradation by heme oxygenase (HO) is a common protective mechanism, and a gene for HO exists in the An. gambiae genome HO (AgHO), although it has yet to be functionally examined. Here, we have cloned and expressed An. gambiae HO (AgHO) in E. coli. Purified recombinant AgHO bound hemin stoichiometrically to form a hemin-enzyme complex similar to other HOs, with a K D of 3.9 ± 0.6 μM; comparable to mammalian and bacterial HOs, but 7-fold lower than that of Drosophila melanogaster HO. AgHO also degraded hemin to biliverdin and released CO and iron in the presence of NADPH cytochrome P450 oxidoreductase (CPR). Optimal AgHO activity was observed at 27.5 °C and pH 7.5. To investigate effects of AgHO inhibition, adult female A. gambiae were fed heme analogues Sn- and Zn-protoporphyrins (SnPP and ZnPP), known to inhibit HO. These led to a dose dependent decrease in oviposition. Cu-protoporphyrin (CuPP), which does not inhibit HO had no effect. These results demonstrate that AgHO is a catalytically active HO and that it may play a key role in egg production in mosquitoes. It also presents a potential target for the development of compounds aimed at sterilising mosquitoes for vector control. Copyright © 2018 Elsevier Ltd. All rights reserved.

A genome-wide screening of BEL-Pao like retrotransposons in Anopheles gambiae by the LTR_STRUC program.

PubMed

Marsano, Renè Massimiliano; Caizzi, Ruggiero

2005-09-12

The advanced status of assembly of the nematoceran Anopheles gambiae genomic sequence allowed us to perform a wide genome analysis to looking at the presence of Long Terminal Repeats (LTRs) in the range of 10 kb by means of the LTR_STRUC tool. More than three hundred sequences were retrieved and 210 were treated as putative complete retrotransposons that were individually analysed with respect to known retrotransposons of A. gambiae and D. melanogaster. The results show that the vast majority of the retrotransposons analysed belong to the Ty3/gypsy class and only 8% to the Ty1/copia class. In addition, phylogenetic analysis allowed us to characterize in more detail the relationship of a large BEL-Pao lineage in which a single family was shown to harbour an additional env gene.

Intra-instar larval cannibalism in Anopheles gambiae (s.s.) and Anopheles stephensi (Diptera: Culicidae).

PubMed

Porretta, Daniele; Mastrantonio, Valentina; Crasta, Graziano; Bellini, Romeo; Comandatore, Francesco; Rossi, Paolo; Favia, Guido; Bandi, Claudio; Urbanelli, Sandra

2016-11-02

Cannibalism has been observed in a wide range of animal taxa and its importance in persistence and stability of populations has been documented. In anopheline malaria vectors the inter-instar cannibalism between fourth- and first-instar larvae (L4-L1) has been shown in several species, while intra-instar cannibalism remains poorly investigated. In this study we tested the occurrence of intra-instar cannibalism within larvae of second-, third- and fourth-instar (L2, L3 and L4) of Anopheles gambiae (s.s.) and An. stephensi. Experiments were set up under laboratory conditions and the effects of larval density, duration of the contact period among larvae and the presence of an older larva (i.e. a potential cannibal of bigger size) on cannibalism rate were analysed. Cannibalism was assessed by computing the number of missing larvae after 24 and 48 h from the beginning of the experiments and further documented by records with a GoPro videocamera. Intra-instar cannibalism was observed in all larval instars of both species with higher frequency in An. gambiae (s.s.) than in An. stephensi. In both species the total number of cannibalistic events increased from 0-24 to 0-48 h. The density affected the cannibalism rate, but its effect was related to the larval instar and to the presence of older larvae. Interestingly, the lower cannibalism rate between L4 larvae was observed at the highest density and the cannibalism rate between L3 larvae decreased when one L4 was added. The present study provides experimental evidence of intra-instar cannibalism in the malaria vectors An. gambiae (s.s.) and An. stephensi and highlights the possible occurrence of complex interactions between all larval instars potentially present in the breeding sites. We hypothesize that the high density and the presence of a potential cannibal of bigger size could affect the readiness to attack conspecifics, resulting into low risk larval behavior and lower cannibalism rate. The understanding of

Polymorphism at the defensin gene in the Anopheles gambiae complex: testing different selection hypotheses

PubMed Central

Simard, Frédéric; Licht, Monica; Besansky, Nora J.; Lehmann, Tovi

2007-01-01

Genetic variation in defensin, a gene encoding a major effector molecule of insects immune response was analyzed within and between populations of three members of the Anopheles gambiae complex. The species selected included the two anthropophilic species, An. gambiae and An. arabiensis and the most zoophilic species of the complex, An. quadriannulatus. The first species was represented by four populations spanning its extreme genetic and geographical ranges, whereas each of the other two species was represented by a single population. We found (i) reduced overall polymorphism in the mature peptide region and in the total coding region, together with specific reductions in rare and moderately frequent mutations (sites) in the coding region compared with non coding regions, (ii) markedly reduced rate of nonsynonymous diversity compared with synonymous variation in the mature peptide and virtually identical mature peptide across the three species, and (iii) increased divergence between species in the mature peptide together with reduced differentiation between populations of An. gambiae in the same DNA region. These patterns suggest a strong purifying selection on the mature peptide and probably the whole coding region. Because An. quadriannulatus is not exposed to human pathogens, identical mature peptide and similar pattern of polymorphism across species implies that human pathogens played no role as selective agents on this peptide. PMID:17161659

Serine protease-related proteins in the malaria mosquito, Anopheles gambiae.

PubMed

Cao, Xiaolong; Gulati, Mansi; Jiang, Haobo

2017-09-01

Insect serine proteases (SPs) and serine protease homologs (SPHs) participate in digestion, defense, development, and other physiological processes. In mosquitoes, some clip-domain SPs and SPHs (i.e. CLIPs) have been investigated for possible roles in antiparasitic responses. In a recent test aimed at improving quality of gene models in the Anopheles gambiae genome using RNA-seq data, we observed various discrepancies between gene models in AgamP4.5 and corresponding sequences selected from those modeled by Cufflinks, Trinity and Bridger. Here we report a comparative analysis of the 337 SP-related proteins in A. gambiae by examining their domain structures, sequence diversity, chromosomal locations, and expression patterns. One hundred and ten CLIPs contain 1 to 5 clip domains in addition to their protease domains (PDs) or non-catalytic, protease-like domains (PLDs). They are divided into five subgroups: CLIPAs (22) are clip 1-5 -PLD; CLIPBs (29), CLIPCs (12) and CLIPDs (14) are mainly clip-PD; most CLIPEs (33) have a domain structure of PD/PLD-PLD-clip-PLD 0-1 . While expression of the CLIP genes in group-1 is generally low and detected in various tissue- and stage-specific RNA-seq libraries, some putative GPs/GPHs (i.e. single domain gut SPs/SPHs) in group-2 are highly expressed in midgut, whole larva or whole adult libraries. In comparison, 46 SPs, 26 SPHs, and 37 multi-domain SPs/SPHs (i.e. PD/PLD-PLD ≥1 ) in group-3 do not seem to be specifically expressed in digestive tract. There are 16 SPs and 2 SPH containing other types of putative regulatory domains (e.g. LDLa, CUB, Gd). Of the 337 SP and SPH genes, 159 were sorted into 46 groups (2-8 members/group) based on similar phylogenetic tree position, chromosomal location, and expression profile. This information and analysis, including improved gene models and protein sequences, constitute a solid foundation for functional analysis of the SP-related proteins in A. gambiae. Copyright © 2017 Elsevier Ltd

Co-occurrence and distribution of East (L1014S) and West (L1014F) African knock-down resistance in Anopheles gambiae sensu lato population of Tanzania

PubMed Central

Kabula, Bilali; Kisinza, William; Tungu, Patrick; Ndege, Chacha; Batengana, Benard; Kollo, Douglas; Malima, Robert; Kafuko, Jessica; Mohamed, Mahdi; Magesa, Stephen

2014-01-01

Objective Insecticide resistance molecular markers can provide sensitive indicators of resistance development in Anopheles vector populations. Assaying these makers is of paramount importance in the resistance monitoring programme. We investigated the presence and distribution of knock-down resistance (kdr) mutations in Anopheles gambiae s.l. in Tanzania. Methods Indoor-resting Anopheles mosquitoes were collected from 10 sites and tested for insecticide resistance using the standard WHO protocol. Polymerase chain reaction-based molecular diagnostics were used to genotype mosquitoes and detect kdr mutations. Results The An. gambiae tested were resistance to lambdacyhalothrin in Muheza, Arumeru and Muleba. Out of 350 An. gambiae s.l. genotyped, 35% were An. gambiae s.s. and 65% An. arabiensis. L1014S and L1014F mutations were detected in both An. gambiae s.s. and An. arabiensis. L1014S point mutation was found at the allelic frequency of 4–33%, while L1014F was at the allelic frequency 6–41%. The L1014S mutation was much associated with An. gambiae s.s. (χ2 = 23.41; P < 0.0001) and L1014F associated with An. arabiensis (χ2 = 11.21; P = 0.0008). The occurrence of the L1014S allele was significantly associated with lambdacyhalothrin resistance mosquitoes (Fisher exact P < 0.001). Conclusion The observed co-occurrence of L1014S and L1014F mutations coupled with reports of insecticide resistance in the country suggest that pyrethroid resistance is becoming a widespread phenomenon among our malaria vector populations. The presence of L1014F mutation in this East African mosquito population indicates the spreading of this gene across Africa. The potential operational implications of these findings on malaria control need further exploration. Objectif Les marqueurs moléculaires de la résistance aux insecticides peuvent fournir des indicateurs sensibles du développement de la résistance dans les populations de vecteurs Anopheles. Le test de ces

Comparative larvicidal toxicities of three ecdysone agonists on the mosquitoes Aedes aegypti, Culex quinquefasciatus, and Anopheles gambiae.

PubMed

Beckage, Nancy E; Marion, Ken M; Walton, William E; Wirth, Margaret C; Tan, Frances F

2004-11-01

Ecdysone agonists are hormonally active insect growth regulators that disrupt development of pest insects and have potential for development as insecticides. Their effects have been particularly well-studied in Lepidoptera and Coleoptera, but significantly less is known about their effects on dipterans, particularly aquatic species. The potency of three ecdysone agonists on larvae of 3 mosquito species, Aedes aegypti, Anopheles gambiae, and Culex quinquefasciatus, was examined. Anopheles gambiae was the most susceptible species and Ae. aegypti was the most resistant species to the effects of the three compounds tested. Potency, in descending order, was RH-2485 > RH-5992 > RH-5849. Dose-response relationships were determined for the three agonists; RH-2485 was found to be the most effective endocrine disruptor against all three species. The observed biological effects of these compounds were similar to those reported for other insects, and mosquitoes initiated molting and apolysis but did not complete a molt. In some cases, mosquito larvae synthesized a new cuticle that appeared to be normally sclerotized but the larvae failed to ecdyse and shed the exuvium. These compounds may prove to be valuable insect growth regulators for control of mosquitoes to decrease the frequency of pathogen transmission to humans. Prospects for using these compounds to control mosquitoes in the field are discussed, along with possible impacts on non-target arthropods in mosquito habitats. (c) 2004 Wiley-Liss, Inc.

Odorant-binding protein-based identification of natural spatial repellents for the African malaria mosquito Anopheles gambiae.

PubMed

Kröber, Thomas; Koussis, Konstantinos; Bourquin, Martine; Tsitoura, Panagiota; Konstantopoulou, Maria; Awolola, Taiwo Sam; Dani, Francesca R; Qiao, Huili; Pelosi, Paolo; Iatrou, Kostas; Guerin, Patrick M

2018-05-01

There is increasing interest in the development of effective mosquito repellents of natural origin to reduce transmission of diseases such as malaria and yellow fever. To achieve this we have employed an in vitro competition assay involving odorant-binding proteins (OBPs) of the malaria mosquito, Anopheles gambiae, with a predominantly female expression bias to identify plant essential oils (EOs) containing bioactive compounds that target mosquito olfactory function. EOs and their fractions capable of binding to such OBPs displayed repellence against female mosquitoes in a laboratory repellent assay. Repellent EOs were subjected to gas chromatographic analysis linked to antennogram (EAG) recordings from female A. gambiae to identify the biologically active constituents. Among these compounds cumin alcohol, carvacrol, ethyl cinnamate and butyl cinnamate proved as effective as DEET at an equivalent dose in the repellent assay, and combinations of carvacrol with either butyl cinnamate or cumin alcohol proved to be significantly more effective than DEET in the assay. When tested as spatial repellents in experimental shelters housing sleeping humans in northern Nigeria a binary mixture of carvacrol plus cumin alcohol caused mosquitoes to leave shelters in significantly higher numbers to those induced by DEET in female Anopheles spp. and in numbers equivalent to that of DEET in Culex spp. mosquitoes. These findings indicate an approach for the identification of biologically active molecules of natural origin serving as repellents for mosquitoes. Copyright © 2018 Elsevier Ltd. All rights reserved.

Genome-Wide Patterns of Polymorphism in an Inbred Line of the African Malaria Mosquito Anopheles gambiae

PubMed Central

Turissini, David A.; Gamez, Stephanie; White, Bradley J.

2014-01-01

Anopheles gambiae is a major mosquito vector of malaria in Africa. Although increased use of insecticide-based vector control tools has decreased malaria transmission, elimination is likely to require novel genetic control strategies. It can be argued that the absence of an A. gambiae inbred line has slowed progress toward genetic vector control. In order to empower genetic studies and enable precise and reproducible experimentation, we set out to create an inbred line of this species. We found that amenability to inbreeding varied between populations of A. gambiae. After full-sib inbreeding for ten generations, we genotyped 112 individuals—56 saved prior to inbreeding and 56 collected after inbreeding—at a genome-wide panel of single nucleotide polymorphisms (SNPs). Although inbreeding dramatically reduced diversity across much of the genome, we discovered numerous, discrete genomic blocks that maintained high heterozygosity. For one large genomic region, we were able to definitively show that high diversity is due to the persistent polymorphism of a chromosomal inversion. Inbred lines in other eukaryotes often exhibit a qualitatively similar retention of polymorphism when typed at a small number of markers. Our whole-genome SNP data provide the first strong, empirical evidence supporting associative overdominance as the mechanism maintaining higher than expected diversity in inbred lines. Although creation of A. gambiae lines devoid of nearly all polymorphism may not be feasible, our results provide critical insights into how more fully isogenic lines can be created. PMID:25377942

A simplified high-throughput method for pyrethroid knock-down resistance (kdr) detection in Anopheles gambiae

PubMed Central

Lynd, Amy; Ranson, Hilary; McCall, P J; Randle, Nadine P; Black, William C; Walker, Edward D; Donnelly, Martin J

2005-01-01

Background A single base pair mutation in the sodium channel confers knock-down resistance to pyrethroids in many insect species. Its occurrence in Anopheles mosquitoes may have important implications for malaria vector control especially considering the current trend for large scale pyrethroid-treated bednet programmes. Screening Anopheles gambiae populations for the kdr mutation has become one of the mainstays of programmes that monitor the development of insecticide resistance. The screening is commonly performed using a multiplex Polymerase Chain Reaction (PCR) which, since it is reliant on a single nucleotide polymorphism, can be unreliable. Here we present a reliable and potentially high throughput method for screening An. gambiae for the kdr mutation. Methods A Hot Ligation Oligonucleotide Assay (HOLA) was developed to detect both the East and West African kdr alleles in the homozygous and heterozygous states, and was optimized for use in low-tech developing world laboratories. Results from the HOLA were compared to results from the multiplex PCR for field and laboratory mosquito specimens to provide verification of the robustness and sensitivity of the technique. Results and Discussion The HOLA assay, developed for detection of the kdr mutation, gives a bright blue colouration for a positive result whilst negative reactions remain colourless. The results are apparent within a few minutes of adding the final substrate and can be scored by eye. Heterozygotes are scored when a sample gives a positive reaction to the susceptible probe and the kdr probe. The technique uses only basic laboratory equipment and skills and can be carried out by anyone familiar with the Enzyme-linked immunosorbent assay (ELISA) technique. A comparison to the multiplex PCR method showed that the HOLA assay was more reliable, and scoring of the plates was less ambiguous. Conclusion The method is capable of detecting both the East and West African kdr alleles in the homozygous and

Discriminative feeding behaviour of Anopheles gambiae s.s. on endemic plants in western Kenya

PubMed Central

Manda, H.; Gouagna, L. C.; Nyandat, E.; Kabiru, E. W.; Jackson, R. R.; Foster, W. A.; Githure, J. I.; Beier, J. C.; Hassanali, A.

2009-01-01

Anopheles gambiae Giles s.s. (Diptera: Culicidae) is known to feed on plant sugars, but this is the first experimental study to consider whether it discriminates between plant species. Thirteen perennial plant species were selected on the basis of their local availability within the vicinity of human dwellings and larval habitats of An. gambiae s.s. in western Kenya. Groups of 100 or 200 mosquitoes were released into cages either with a cutting of one plant type at a time (single-plant assay) or with cuttings of all 13 plants simultaneously (choice assay), respectively, and left overnight. In the choice assay, direct observations of the percentages of mosquitoes perching or feeding on each plant were recorded over four 1-h periods each night. For both types of assay, mosquitoes were recaptured and the percentage that had fed on plants was assessed by testing them individually for the presence of fructose. To identify which plants the choice-assay mosquitoes had fed on, gas chromatography (GC) profiles of samples of mosquito homogenates were compared with GC profiles of extracts from relevant parts of each plant. Four of the plants that were observed to have been fed on most frequently in the choice assay (Parthenium hysterophorus L., Tecoma stans L., Ricinus communis L., and Senna didymobotrya Fresen) were also shown to have been ingested most often by mosquitoes in both types of assay, suggesting that An. gambiae is differentially responsive to this range of plants, regardless of whether the plants were presented singly or mixed together. Significantly more females than males fed on plants, with the exception of P. hysterophorus L., one of the plants most frequently fed on. For most plant species (ten of 13), GC profiles indicated that An. gambiae obtained sugars primarily from flowers. The exceptions were P. hysterophorus L., Lantana camara L. and R. communis L., on which An. gambiae fed more often from leaves and stems than from flowers. PMID:17373953

Larval and adult environmental temperatures influence the adult reproductive traits of Anopheles gambiae s.s.

PubMed

Christiansen-Jucht, Céline D; Parham, Paul E; Saddler, Adam; Koella, Jacob C; Basáñez, María-Gloria

2015-09-17

Anopheles mosquito life-history parameters and population dynamics strongly influence malaria transmission, and environmental factors, particularly temperature, strongly affect these parameters. There are currently some studies on how temperature affects Anopheles gambiae s.s. survival but very few exist examining other life-history traits. We investigate here the effect of temperature on population dynamics parameters. Anopheles gambiae s.s. immatures were reared individually at 23 ± 1 °C, 27 ± 1 °C, 31 ± 1 °C, and 35 ± 1 °C, and adults were held at their larval temperature or at one of the other temperatures. Larvae were checked every 24 h for development to the next stage and measured for size; wing length was measured as a proxy for adult size. Females were blood fed three times, and the number of females feeding and laying eggs was counted. The numbers of eggs and percentage of eggs hatched were recorded. Increasing temperatures during the larval stages resulted in significantly smaller larvae (p = 0.005) and smaller adults (p < 0.001). Adult temperature had no effect on the time to egg laying, and the larval temperature of adults only affected the incubation period of the first egg batch. Temperature influenced the time to hatching of eggs, as well as the time to development at every stage. The number of eggs laid was highest when adults were kept at 27 °C, and lowest at 31 °C, and higher adult temperatures decreased the proportion of eggs hatching after the second and third blood meal. Higher adult temperatures significantly decreased the probability of blood feeding, but the larval temperature of adults had no influence on the probability of taking a blood meal. Differences were observed between the first, second, and third blood meal in the times to egg laying and hatching, number of eggs laid, and probabilities of feeding and laying eggs. Our study shows that environmental temperature during the larval stages as

Orientation of Anopheles gambiae (Diptera: Culicidae) to Plant-Host Volatiles in a Novel Diffusion-Cage Olfactometer

PubMed Central

Otienoburu, Philip E.; Nikbakhtzadeh, Mahmood R.; Foster, Woodbridge A.

2016-01-01

A novel diffusion-cage olfactometer tested the responses of Anopheles gambiae Giles to plant volatiles. Green-leaf volatiles are often released from cut or injured plant tissue and may alter the headspace of plants used in olfactometer assays. The diffusion-cage olfactometer is designed for use with whole, intact plants, hence giving a more realistic behavioral assay. Its simple plastic construction, ease of assembly, and accommodation to whole plants makes it a useful tool for measuring mosquito orientation to plant volatiles within large enclosures. We compared its performance to that of the more commonly used T-tube wind-tunnel olfactometer, by testing the orientation of mosquitoes to volatiles of a few prevalent plants of eastern Africa reportedly utilized by An. gambiae for sugar: Parthenium hysterophorus (Asteraceae), Ricinus communis (Euphorbiaceae), Lantana camara (Verbenaceae), and Senna occidentalis (Fabaceae). Results indicate that the diffusion-cage olfactometer is an effective alternative to conventional wind-tunnel olfactometers, to test mosquito orientation to plant volatiles under seminatural conditions. PMID:26502752

The current insecticide resistance status of Anopheles gambiae (s.l.) (Culicidae) in rural and urban areas of Bouaké, Côte d'Ivoire.

PubMed

Zoh, Dounin Danielle; Ahoua Alou, Ludovic Phamien; Toure, Mahama; Pennetier, Cédric; Camara, Soromane; Traore, Dipomin François; Koffi, Alphonsine Amanan; Adja, Akré Maurice; Yapi, Ahoua; Chandre, Fabrice

2018-03-02

Several studies were carried out in experimental hut station in areas surrounding the city of Bouaké, after the crisis in Côte d'Ivoire. They reported increasing resistance levels to insecticide for malaria transmiting mosquitoes. The present work aims to evaluate the current resistance level of An. gambiae (s.l.) in rural and urban areas in the city of Bouaké. Larvae of Anopheles gambiae (s.l.) were collected from five different study sites and reared to adult stages. The resistance status was assessed using the WHO bioassay test kits for adult mosquitoes, with eight insecticides belonging to pyrethroids, organochlorines, carbamates and organophosphates classes. Molecular assays were performed to identify the molecular forms of An. gambiae (s.l.), the L1014F kdr and the ace-1R alleles in individual mosquitoes. The synergist PBO was used to investigate the role of enzymes in resistance. Biochemical assays were performed to detect potential increased activities in mixed function oxidase (MFO) levels, non-specific esterases (NSE) and glutathione S-transferases (GST). High resistance levels to pyrethroids, organochlorines, and carbamates were observed in Anopheles gambiae (s.l.) from Bouaké. Mortalities ranged between 0 and 73% for the eight tested insecticides. The pre-exposure to PBO restored full or partial susceptibility to pyrethroids in the different sites. The same trend was observed with the carbamates in five sites, but to a lesser extent. With DDT, pre-exposure to PBO did not increase the mortality rate of An. gambiae (s.l.) from the same sites. Tolerance to organophosphates was observed. An increased activity of NSE and higher level of MFO were found compared to the Kisumu susceptible reference strain. Two molecular forms, S form [(An. gambiae (s.s)] and M form (An. coluzzi) were identified. The kdr allele frequencies vary from 85.9 to 99.8% for An. gambiae (s.s.) and from 81.7 to 99.6% for An. coluzzii. The ace-1R frequencies vary between 25.6 and 38

Larvicidal effects of a neem (Azadirachta indica) oil formulation on the malaria vector Anopheles gambiae.

PubMed

Okumu, Fredros O; Knols, Bart G J; Fillinger, Ulrike

2007-05-22

Larviciding is a key strategy used in many vector control programmes around the world. Costs could be reduced if larvicides could be manufactured locally. The potential of natural products as larvicides against the main African malaria vector, Anopheles gambiae s.s was evaluated. To assess the larvicidal efficacy of a neem (Azadirachta indica) oil formulation (azadirachtin content of 0.03% w/v) on An. gambiae s.s., larvae were exposed as third and fourth instars to a normal diet supplemented with the neem oil formulations in different concentrations. A control group of larvae was exposed to a corn oil formulation in similar concentrations. Neem oil had an LC50 value of 11 ppm after 8 days, which was nearly five times more toxic than the corn oil formulation. Adult emergence was inhibited by 50% at a concentration of 6 ppm. Significant reductions on growth indices and pupation, besides prolonged larval periods, were observed at neem oil concentrations above 8 ppm. The corn oil formulation, in contrast, produced no growth disruption within the tested range of concentrations. Neem oil has good larvicidal properties for An. gambiae s.s. and suppresses successful adult emergence at very low concentrations. Considering the wide distribution and availability of this tree and its products along the East African coast, this may prove a readily available and cheap alternative to conventional larvicides.

Exposure to disinfectants (soap or hydrogen peroxide) increases tolerance to permethrin in Anopheles gambiae populations from the city of Yaoundé, Cameroon.

PubMed

Antonio-Nkondjio, Christophe; Youmsi-Goupeyou, Marlene; Kopya, Edmond; Tene-Fossog, Billy; Njiokou, Flobert; Costantini, Carlo; Awono-Ambene, Parfait

2014-08-03

The rapid expansion of insecticide resistance is limiting the efficiency of malaria vector control interventions. However, current knowledge of factors inducing pyrethroid resistance remains incomplete. In the present study, the role of selection at the larval stage by disinfectants, such as soap and hydrogen peroxide (H2O2), on adult mosquito resistance to permethrin was investigated. Field Anopheles gambiae sensu lato larvae, were exposed to variable concentrations of soap and H2O2. Larvae surviving to acute toxicity assays after 24 hours were reared to the adult stage and exposed to permethrin. The susceptibility level of adults was compared to the untreated control group. The effect of soap or hydrogen peroxide selection on the length of larval development and emergence rate was assessed. Larval bioassays analysis showed a more acute effect of hydrogen peroxide on mosquito larvae compared to soap. The regression lines describing the dose mortality profile showed higher mean and variance to hydrogen peroxide than to soap. The duration of larval development (<5 days) and adults emergence rates (1 to 77%) were shorter and lower compare to control. Anopheles gambiae s.l. larvae surviving to selection with either soap or hydrogen peroxide or both, produced adults who were up to eight-times more resistant to permethrin than mosquitoes from the untreated control group. The present study shows that selective pressure exerted by non-insecticidal compounds such as soap and hydrogen peroxide affect An. gambiae s.l. tolerance to pyrethroids. This requires further studies with regard to the adaptation of An. gambiae s.l. to polluted habitats across sub-Saharan Africa cities.

Anopheles salivary gland proteomes from major malaria vectors

PubMed Central

2012-01-01

Background Antibody responses against Anopheles salivary proteins can indicate individual exposure to bites of malaria vectors. The extent to which these salivary proteins are species-specific is not entirely resolved. Thus, a better knowledge of the diversity among salivary protein repertoires from various malaria vector species is necessary to select relevant genus-, subgenus- and/or species-specific salivary antigens. Such antigens could be used for quantitative (mosquito density) and qualitative (mosquito species) immunological evaluation of malaria vectors/host contact. In this study, salivary gland protein repertoires (sialomes) from several Anopheles species were compared using in silico analysis and proteomics. The antigenic diversity of salivary gland proteins among different Anopheles species was also examined. Results In silico analysis of secreted salivary gland protein sequences retrieved from an NCBInr database of six Anopheles species belonging to the Cellia subgenus (An. gambiae, An. arabiensis, An. stephensi and An. funestus) and Nyssorhynchus subgenus (An. albimanus and An. darlingi) displayed a higher degree of similarity compared to salivary proteins from closely related Anopheles species. Additionally, computational hierarchical clustering allowed identification of genus-, subgenus- and species-specific salivary proteins. Proteomic and immunoblot analyses performed on salivary gland extracts from four Anopheles species (An. gambiae, An. arabiensis, An. stephensi and An. albimanus) indicated that heterogeneity of the salivary proteome and antigenic proteins was lower among closely related anopheline species and increased with phylogenetic distance. Conclusion This is the first report on the diversity of the salivary protein repertoire among species from the Anopheles genus at the protein level. This work demonstrates that a molecular diversity is exhibited among salivary proteins from closely related species despite their common pharmacological

Sugar-sensitive neurone responses and sugar feeding preferences influence lifespan and biting behaviours of the Afrotropical malaria mosquito, Anopheles gambiae.

PubMed

Kessler, Sébastien; Vlimant, Michèle; Guerin, Patrick M

2015-03-01

Floral nectar is the main source of carbohydrates for many insects including mosquitoes. Nonetheless, the physiological mechanisms underlying feeding on carbohydrates by the Afrotropical malaria mosquito Anopheles gambiae remain poorly understood. Here, we tested whether sugar sensitivity and sugar feeding preferences correlate with longevity in A. gambiae. We also tested whether feeding females on different sugar diets influences their biting behaviours. Electrophysiological recordings show that sugar neurones on the labella of females are most sensitive to sucrose, mixtures of glucose and fructose, and to melezitose; other sugars tested, including glucose and fructose presented alone, only weakly activate these taste neurones. Mosquitoes survive longest on sucrose, the most preferred sugar. Whereas feeding on a mixture of glucose and fructose is preferred over fructose or glucose alone, fructose supports higher longevity than either glucose or the mixture of the two hexoses. Females that had previously fed on glucose show a stronger biting response than those fed on sucrose, perhaps in an effort to compensate for their lower energetic state. These findings contribute to our understanding of the physiological basis of sugar feeding in A. gambiae and indicate how the sugar diet can affect laboratory-reared A. gambiae biting behaviours.

Cytochrome P450 associated with insecticide resistance catalyzes cuticular hydrocarbon production in Anopheles gambiae.

PubMed

Balabanidou, Vasileia; Kampouraki, Anastasia; MacLean, Marina; Blomquist, Gary J; Tittiger, Claus; Juárez, M Patricia; Mijailovsky, Sergio J; Chalepakis, George; Anthousi, Amalia; Lynd, Amy; Antoine, Sanou; Hemingway, Janet; Ranson, Hilary; Lycett, Gareth J; Vontas, John

2016-08-16

The role of cuticle changes in insecticide resistance in the major malaria vector Anopheles gambiae was assessed. The rate of internalization of (14)C deltamethrin was significantly slower in a resistant strain than in a susceptible strain. Topical application of an acetone insecticide formulation to circumvent lipid-based uptake barriers decreased the resistance ratio by ∼50%. Cuticle analysis by electron microscopy and characterization of lipid extracts indicated that resistant mosquitoes had a thicker epicuticular layer and a significant increase in cuticular hydrocarbon (CHC) content (∼29%). However, the CHC profile and relative distribution were similar in resistant and susceptible insects. The cellular localization and in vitro activity of two P450 enzymes, CYP4G16 and CYP4G17, whose genes are frequently overexpressed in resistant Anopheles mosquitoes, were analyzed. These enzymes are potential orthologs of the CYP4G1/2 enzymes that catalyze the final step of CHC biosynthesis in Drosophila and Musca domestica, respectively. Immunostaining indicated that both CYP4G16 and CYP4G17 are highly abundant in oenocytes, the insect cell type thought to secrete hydrocarbons. However, an intriguing difference was indicated; CYP4G17 occurs throughout the cell, as expected for a microsomal P450, but CYP4G16 localizes to the periphery of the cell and lies on the cytoplasmic side of the cell membrane, a unique position for a P450 enzyme. CYP4G16 and CYP4G17 were functionally expressed in insect cells. CYP4G16 produced hydrocarbons from a C18 aldehyde substrate and thus has bona fide decarbonylase activity similar to that of dmCYP4G1/2. The data support the hypothesis that the coevolution of multiple mechanisms, including cuticular barriers, has occurred in highly pyrethroid-resistant An gambiae.

Cytochrome P450 associated with insecticide resistance catalyzes cuticular hydrocarbon production in Anopheles gambiae

PubMed Central

Balabanidou, Vasileia; Kampouraki, Anastasia; MacLean, Marina; Blomquist, Gary J.; Tittiger, Claus; Juárez, M. Patricia; Mijailovsky, Sergio J.; Chalepakis, George; Anthousi, Amalia; Lynd, Amy; Antoine, Sanou; Hemingway, Janet; Ranson, Hilary; Lycett, Gareth J.; Vontas, John

2016-01-01

The role of cuticle changes in insecticide resistance in the major malaria vector Anopheles gambiae was assessed. The rate of internalization of 14C deltamethrin was significantly slower in a resistant strain than in a susceptible strain. Topical application of an acetone insecticide formulation to circumvent lipid-based uptake barriers decreased the resistance ratio by ∼50%. Cuticle analysis by electron microscopy and characterization of lipid extracts indicated that resistant mosquitoes had a thicker epicuticular layer and a significant increase in cuticular hydrocarbon (CHC) content (∼29%). However, the CHC profile and relative distribution were similar in resistant and susceptible insects. The cellular localization and in vitro activity of two P450 enzymes, CYP4G16 and CYP4G17, whose genes are frequently overexpressed in resistant Anopheles mosquitoes, were analyzed. These enzymes are potential orthologs of the CYP4G1/2 enzymes that catalyze the final step of CHC biosynthesis in Drosophila and Musca domestica, respectively. Immunostaining indicated that both CYP4G16 and CYP4G17 are highly abundant in oenocytes, the insect cell type thought to secrete hydrocarbons. However, an intriguing difference was indicated; CYP4G17 occurs throughout the cell, as expected for a microsomal P450, but CYP4G16 localizes to the periphery of the cell and lies on the cytoplasmic side of the cell membrane, a unique position for a P450 enzyme. CYP4G16 and CYP4G17 were functionally expressed in insect cells. CYP4G16 produced hydrocarbons from a C18 aldehyde substrate and thus has bona fide decarbonylase activity similar to that of dmCYP4G1/2. The data support the hypothesis that the coevolution of multiple mechanisms, including cuticular barriers, has occurred in highly pyrethroid-resistant An. gambiae. PMID:27439866

First report of the East African kdr mutation in an Anopheles gambiae mosquito in Côte d'Ivoire.

PubMed

Chouaïbou, Mouhamadou; Kouadio, Fodjo Behi; Tia, Emmanuel; Djogbenou, Luc

2017-02-09

Background . The intensive use of insecticides in public health and agriculture has led to the development of insecticide resistances in malaria vectors across sub-Saharan Africa countries in the last two decades. The kdr target site point mutation which is among the best characterised resistance mechanisms seems to be changing its distribution patterns on the African continent. The 1014F  kdr mutation originally described only in West Africa is spreading to East Africa while the 1014S  kdr mutation originally described in East Africa, is spreading to West and Central Africa. However, the East- kdr mutation has not been reported in Côte d'Ivoire so far. Methods . Immature stages of Anopheles gambiae s.l. were collected from breeding sites at the outskirts of Yamoussoukro, Côte d'Ivoire. Emerging 3-5 day old adult female mosquitoes were tested for susceptibility to deltamethrin 0.05%, malathion 5%, bendiocarb 1% and dichlorodiphenyltrichloroethane (DDT) 4% according to WHO standard procedures. A total of 50 An. gambiae s.l. specimens were drawn at random for DNA extraction and identification down to the species level. A subsample of 30 mosquitoes was tested for the East-African kdr mutation using a Taqman assay. Results . The tested mosquito population appeared to be strongly resistant to deltamethrin (1.03% mortality), bendiocarb (38.46% mortality) and DDT (0% mortality) with probable resistance observed for malathion (92.47%). Among the 41 mosquitoes that were successfully characterized, An. coluzzii was predominant (68.3%) followed by An. gambiae   s.s. (19.5%) and a few hybrids (7.3%). Out of 30 specimens genotyped for East- kdr , a single hybrid mosquito appeared to be heterozygous for the mutation. Conclusion . The present study revealed the presence of the East- kdr mutation in Côte d'Ivoire for the first time in An. gambiae and highlights the urgent need to start monitoring the allele and genotype frequencies.

Remote sensing and environment in the study of the malaria vector Anopheles gambiae in Mali

NASA Astrophysics Data System (ADS)

Rian, Sigrid Katrine Eivindsdatter

The malaria mosquito Anopheles gambiae is the most important vector for the most devastating form of human malaria, the parasite Plasmodium falciparum. In-depth knowledge of the vector's history and environmental preferences is essential in the pursuit of new malaria mitigation strategies. Research was conducted in Mali across a range of habitats occupied by the vector, focusing on three identified chromosomal forms in the mosquito complex. The development of a 500-m landcover classification map was carried out using MODIS satellite imagery and extensive ground survey. The resulting product has the highest resolution and is the most up-to-date and most extensively ground-surveyed among land-cover maps for the study region. The new landcover classification product is a useful tool in the mapping of the varying ecological preferences of the different An. gambiae chromosomal forms. Climate and vegetation characteristics and their relationship to chromosomal forms were investigated further along a Southwest-Northeast moisture gradient in Mali. This research demonstrates particular ecological preferences of each chromosomal form, and gives a detailed examination of particular vegetation structural and climatological patterns across the study region. A key issue in current research into the population structure of An. gambiae is speciation and evolution in the complex, as an understanding of the mechanisms of change can help in the development of new mitigation strategies. A historical review of the paleoecology, archaeology, and other historical sources intended to shed light on the evolutionary history of the vector is presented. The generally held assumption that the current breed of An. gambiae emerged in the rainforest is called into question and discussed within the framework of paleoenvironment and human expansions in sub-Saharan West Africa.

Larvicidal effect of disinfectant soap on Anopheles gambiae s.s (Diptera: Culicidae) in laboratory and semifield environs

PubMed Central

2014-01-01

Background Mosquito larval control using chemicals and biological agents is of paramount importance in vector population and disease incidence reduction. A commercial synthetic disinfectant soap was evaluated against larvae of Anopheles gambiae s.s. in both laboratory and semi field conditions. Method Five concentrations of commercial synthetic disinfectant soap (0.0001, 0.001, 0.01, 0.1 and 1%) were prepared and evaluated against third instar larvae in laboratory and semi field environments. Mortality was scored at 12, 24, 48, and 72 hrs. Each dosage had 6 replicates, having twenty 3rd instar larvae of An.gambiae s.s. Results In the laboratory phase, all dosages had significantly higher larval mortalities than in controls, while in semi field conditions, the dosages of 0.0001, 0.001 and 0.01% had lower mortalities than laboratory trials. In the comparison between semi field and laboratory trials, only 0.1 and 1% dosage had significant difference with more mortality in semifield conditions. Proportions of larvae that died during mortality monitoring intervals in laboratory and semi field had significant differences only at 12 hrs and 72 hrs. Conclusion The findings of this study have demonstrated that the mortality of larvae caused by commercial synthetic disinfectant soap is worth further studies in open water bodies. More studies are necessary to find out the effect of sunlight on the chemistry of the synthetic disinfectant and other variables in small scale full field trials. PMID:24885903

CRISPR/Cas9 -mediated gene knockout of Anopheles gambiae FREP1 suppresses malaria parasite infection

PubMed Central

Dong, Yuemei; Simões, Maria L.

2018-01-01

Plasmodium relies on numerous agonists during its journey through the mosquito vector, and these agonists represent potent targets for transmission-blocking by either inhibiting or interfering with them pre- or post-transcriptionally. The recently developed CRISPR/Cas9-based genome editing tools for Anopheles mosquitoes provide new and promising opportunities for the study of agonist function and for developing malaria control strategies through gene deletion to achieve complete agonist inactivation. Here we have established a modified CRISPR/Cas9 gene editing procedure for the malaria vector Anopheles gambiae, and studied the effect of inactivating the fibrinogen-related protein 1 (FREP1) gene on the mosquito’s susceptibility to Plasmodium and on mosquito fitness. FREP1 knockout mutants developed into adult mosquitoes that showed profound suppression of infection with both human and rodent malaria parasites at the oocyst and sporozoite stages. FREP1 inactivation, however, resulted in fitness costs including a significantly lower blood-feeding propensity, fecundity and egg hatching rate, a retarded pupation time, and reduced longevity after a blood meal. PMID:29518156

Metabolic and Target-Site Mechanisms Combine to Confer Strong DDT Resistance in Anopheles gambiae

PubMed Central

Mitchell, Sara N.; Rigden, Daniel J.; Dowd, Andrew J.; Lu, Fang; Wilding, Craig S.; Weetman, David; Dadzie, Samuel; Jenkins, Adam M.; Regna, Kimberly; Boko, Pelagie; Djogbenou, Luc; Muskavitch, Marc A. T.; Ranson, Hilary; Paine, Mark J. I.; Mayans, Olga; Donnelly, Martin J.

2014-01-01

The development of resistance to insecticides has become a classic exemplar of evolution occurring within human time scales. In this study we demonstrate how resistance to DDT in the major African malaria vector Anopheles gambiae is a result of both target-site resistance mechanisms that have introgressed between incipient species (the M- and S-molecular forms) and allelic variants in a DDT-detoxifying enzyme. Sequencing of the detoxification enzyme, Gste2, from DDT resistant and susceptible strains of An. gambiae, revealed a non-synonymous polymorphism (I114T), proximal to the DDT binding domain, which segregated with strain phenotype. Recombinant protein expression and DDT metabolism analysis revealed that the proteins from the susceptible strain lost activity at higher DDT concentrations, characteristic of substrate inhibition. The effect of I114T on GSTE2 protein structure was explored through X-ray crystallography. The amino acid exchange in the DDT-resistant strain introduced a hydroxyl group nearby the hydrophobic DDT-binding region. The exchange does not result in structural alterations but is predicted to facilitate local dynamics and enzyme activity. Expression of both wild-type and 114T alleles the allele in Drosophila conferred an increase in DDT tolerance. The 114T mutation was significantly associated with DDT resistance in wild caught M-form populations and acts in concert with target-site mutations in the voltage gated sodium channel (Vgsc-1575Y and Vgsc-1014F) to confer extreme levels of DDT resistance in wild caught An. gambiae. PMID:24675797

Insecticide resistance mechanisms associated with different environments in the malaria vector Anopheles gambiae: a case study in Tanzania.

PubMed

Nkya, Theresia E; Akhouayri, Idir; Poupardin, Rodolphe; Batengana, Bernard; Mosha, Franklin; Magesa, Stephen; Kisinza, William; David, Jean-Philippe

2014-01-25

Resistance of mosquitoes to insecticides is a growing concern in Africa. Since only a few insecticides are used for public health and limited development of new molecules is expected in the next decade, maintaining the efficacy of control programmes mostly relies on resistance management strategies. Developing such strategies requires a deep understanding of factors influencing resistance together with characterizing the mechanisms involved. Among factors likely to influence insecticide resistance in mosquitoes, agriculture and urbanization have been implicated but rarely studied in detail. The present study aimed at comparing insecticide resistance levels and associated mechanisms across multiple Anopheles gambiae sensu lato populations from different environments. Nine populations were sampled in three areas of Tanzania showing contrasting agriculture activity, urbanization and usage of insecticides for vector control. Insecticide resistance levels were measured in larvae and adults through bioassays with deltamethrin, DDT and bendiocarb. The distribution of An. gambiae sub-species and pyrethroid target-site mutations (kdr) were investigated using molecular assays. A microarray approach was used for identifying transcription level variations associated to different environments and insecticide resistance. Elevated resistance levels to deltamethrin and DDT were identified in agriculture and urban areas as compared to the susceptible strain Kisumu. A significant correlation was found between adult deltamethrin resistance and agriculture activity. The subspecies Anopheles arabiensis was predominant with only few An. gambiae sensu stricto identified in the urban area of Dar es Salaam. The L1014S kdr mutation was detected at elevated frequency in An gambiae s.s. in the urban area but remains sporadic in An. arabiensis specimens. Microarrays identified 416 transcripts differentially expressed in any area versus the susceptible reference strain and supported the impact

Olfactory basis of floral preference of the malaria vector Anopheles gambiae (Diptera: Culicidae) among common African plants.

PubMed

Nikbakhtzadeh, Mahmood R; Terbot, John W; Otienoburu, Philip E; Foster, Woodbridge A

2014-12-01

Mosquitoes of both sexes feed on plants to obtain sugar. Nocturnal species probably locate the plants primarily by their volatile semiochemicals that also form the basis for the mosquitoes' innate plant-species preferences. To evaluate these olfactory preferences quantitatively, we used a two-choice wind-tunnel olfactometer to measure the upwind orientation of Anopheles gambiae Giles, an important vector of malaria in equatorial Africa, toward odor plumes produced by nine plant species common where this mosquito occurs. These plants are reported to induce feeding behaviors in An. gambiae and to produce floral or extrafloral nectar. Results presented here demonstrated that the volatiles of S. didymobotrya, P. hysterophorus, S. occidentalis, and L. camara, in descending order of numbers of mosquitoes responding, were all attractive, compared to a control plant species, whereas D. stramonium, R. communis, S. bicapsularis, T. stans, and T. diversifolia were not. As expected, chromatographic analysis of the headspace of attractive plants whose volatiles were captured by stir-bar sorptive extraction revealed a wide range of compounds, primarily terpenoids. Once their bioactivity and attractiveness for An. gambiae, alone and in blends, has been firmly established, some of these semiochemicals may have applications in population sampling and control. © 2014 The Society for Vector Ecology.

Analysis of natural female post-mating responses of Anopheles gambiae and Anopheles coluzzii unravels similarities and differences in their reproductive ecology.

PubMed

Thailayil, Janis; Gabrieli, Paolo; Caputo, Beniamino; Bascuñán, Priscila; South, Adam; Diabate, Abdoulaye; Dabire, Roch; Della Torre, Alessandra; Catteruccia, Flaminia

2018-04-26

Anopheles gambiae and An. coluzzii, the two most important malaria vectors in sub-Saharan Africa, are recently radiated sibling species that are reproductively isolated even in areas of sympatry. In females from these species, sexual transfer of male accessory gland products, including the steroid hormone 20-hydroxyecdysone (20E), induces vast behavioral, physiological, and transcriptional changes that profoundly shape their post-mating ecology, and that may have contributed to the insurgence of post-mating, prezygotic reproductive barriers. As these barriers can be detected by studying transcriptional changes induced by mating, we set out to analyze the post-mating response of An. gambiae and An. coluzzii females captured in natural mating swarms in Burkina Faso. While the molecular pathways shaping short- and long-term mating-induced changes are largely conserved in females from the two species, we unravel significant inter-specific differences that suggest divergent regulation of key reproductive processes such as egg development, processing of seminal secretion, and mating behavior, that may have played a role in reproductive isolation. Interestingly, a number of these changes occur in genes previously shown to be regulated by the sexual transfer of 20E and may be due to divergent utilization of this steroid hormone in the two species.

Visualizing non infectious and infectious Anopheles gambiae blood feedings in naive and saliva-immunized mice.

PubMed

Choumet, Valerie; Attout, Tarik; Chartier, Loïc; Khun, Huot; Sautereau, Jean; Robbe-Vincent, Annie; Brey, Paul; Huerre, Michel; Bain, Odile

2012-01-01

Anopheles gambiae is a major vector of malaria and lymphatic filariasis. The arthropod-host interactions occurring at the skin interface are complex and dynamic. We used a global approach to describe the interaction between the mosquito (infected or uninfected) and the skin of mammals during blood feeding. Intravital video microscopy was used to characterize several features during blood feeding. The deposition and movement of Plasmodium berghei sporozoites in the dermis were also observed. We also used histological techniques to analyze the impact of infected and uninfected feedings on the skin cell response in naive mice. The mouthparts were highly mobile within the skin during the probing phase. Probing time increased with mosquito age, with possible effects on pathogen transmission. Repletion was achieved by capillary feeding. The presence of sporozoites in the salivary glands modified the behavior of the mosquitoes, with infected females tending to probe more than uninfected females (86% versus 44%). A white area around the tip of the proboscis was observed when the mosquitoes fed on blood from the vessels of mice immunized with saliva. Mosquito feedings elicited an acute inflammatory response in naive mice that peaked three hours after the bite. Polynuclear and mast cells were associated with saliva deposits. We describe the first visualization of saliva in the skin by immunohistochemistry (IHC) with antibodies directed against saliva. Both saliva deposits and sporozoites were detected in the skin for up to 18 h after the bite. This study, in which we visualized the probing and engorgement phases of Anopheles gambiae blood meals, provides precise information about the behavior of the insect as a function of its infection status and the presence or absence of anti-saliva antibodies. It also provides insight into the possible consequences of the inflammatory reaction for blood feeding and pathogen transmission.

Salivary gland proteome analysis reveals modulation of anopheline unique proteins in insensitive acetylcholinesterase resistant Anopheles gambiae mosquitoes.

PubMed

Cornelie, Sylvie; Rossignol, Marie; Seveno, Martial; Demettre, Edith; Mouchet, François; Djègbè, Innocent; Marin, Philippe; Chandre, Fabrice; Corbel, Vincent; Remoué, Franck; Mathieu-Daudé, Françoise

2014-01-01

Insensitive acetylcholinesterase resistance due to a mutation in the acetylcholinesterase (ace) encoding ace-1 gene confers cross-resistance to organophosphate and carbamate insecticides in Anopheles gambiae populations from Central and West Africa. This mutation is associated with a strong genetic cost revealed through alterations of some life history traits but little is known about the physiological and behavioural changes in insects bearing the ace-1(R) allele. Comparative analysis of the salivary gland contents between An. gambiae susceptible and ace-1(R) resistant strains was carried out to charaterize factors that could be involved in modifications of blood meal process, trophic behaviour or pathogen interaction in the insecticide-resistant mosquitoes. Differential analysis of the salivary gland protein profiles revealed differences in abundance for several proteins, two of them showing major differences between the two strains. These two proteins identified as saglin and TRIO are salivary gland-1 related proteins, a family unique to anopheline mosquitoes, one of them playing a crucial role in salivary gland invasion by Plasmodium falciparum sporozoites. Differential expression of two other proteins previously identified in the Anopheles sialome was also observed. The differentially regulated proteins are involved in pathogen invasion, blood feeding process, and protection against oxidation, relevant steps in the outcome of malaria infection. Further functional studies and insect behaviour experiments would confirm the impact of the modification of the sialome composition on blood feeding and pathogen transmission abilities of the resistant mosquitoes. The data supports the hypothesis of alterations linked to insecticide resistance in the biology of the primary vector of human malaria in Africa.

Species Identification and Resistance Status of Anopheles gambiae s.l. (Diptera: Culicidae) Mosquitoes in Guinea.

PubMed

Keita, K; Camara, D; Barry, Y; Ossè, R; Wang, L; Sylla, M; Miller, D; Leite, L; Schopp, P; Lawrence, G G; Akogbéto, M; Dotson, E M; Guilavogui, T; Keita, M; Irish, S R

2017-05-01

Insecticide resistance is one of the primary threats to the recent gains in malaria control. This is especially true in Guinea, where long-lasting insecticidal nets are currently the primary vector control intervention. To better inform the national malaria control program on the current status of insecticide resistance in Guinea, resistance bioassays were conducted, using Anopheles gambiae s.l. Giles, in three sites. Molecular analyses were also done on An. gambiae s.l. to determine the species and find whether the target-site mutations kdr and Ace1R were present. Susceptibility tests revealed resistance to DDT and pyrethroids, although mosquitoes were susceptible to deltamethrin in two of the three sites tested. Mosquitoes were susceptible to bendiocarb, except in Kissidougou, Guinea. The kdr-west mutation was widespread and the frequency was 60% or more in all sites. However, the Ace1R mutation was present in low levels. Insecticide susceptibility should continue to be monitored in Guinea to ensure insecticide-based vector control methods remain effective. Published by Oxford University Press on behalf of Entomological Society of America 2017. This work is written by US Government employees and is in the public domain in the US.

Application of a reverse dot blot, DNA-DNA hydridization method to quantify host-feeding tendencies of two sibling species in the Anopheles gambiae complex

PubMed Central

Fritz, Megan L; Miller, James R; Bayoh, M Nabie; Vulule, John M; Landgraf, Jeffrey R; Walker, Edward D

2012-01-01

A DNA-DNA hybridization method, reverse dot blot analysis (RDBA), was used for identification of Anopheles gambiae s.s. and An. arabiensis hosts. Of 299 blood fed and half gravid An. gambiae s.l. collected from Kisian, Kenya, 244 individuals were identifiable to species; 69.5% were An. arabiensis, and 29.5% were An. gambiae s.s. Host identifications with RDBA were comparable to conventional PCR followed by direct sequencing of amplicons of the vertebrate mitochondrial cytochrome B gene. Of the 174 amplicon-producing samples used for comparison of these two methods, 147 were identifiable by direct sequencing, and 139 of these same by RDBA. An. arabiensis blood meals were mostly (>90%) bovine in origin, whereas An. gambiae s.s. fed upon humans > 90% of the time. RDBA detected that 2 of 112 An. arabiensis had blood from more than one host species, whereas PCR and direct sequencing did not. Recent insecticide-treated bednet (ITN) use in Kisian has likely caused the shift in the dominant vector species from An. gambiae s.s. to An. arabiensis. RDBA provides an opportunity to study changes in host-feeding by members of the An. gambiae complex as a response to the broadening distribution of vector control measures targeting host-selection behaviors. PMID:24188164

Comprehensive Genetic Dissection of the Hemocyte Immune Response in the Malaria Mosquito Anopheles gambiae

PubMed Central

Lombardo, Fabrizio; Ghani, Yasmeen; Kafatos, Fotis C.; Christophides, George K.

2013-01-01

Reverse genetics in the mosquito Anopheles gambiae by RNAi mediated gene silencing has led in recent years to an advanced understanding of the mosquito immune response against infections with bacteria and malaria parasites. We developed RNAi screens in An. gambiae hemocyte-like cells using a library of double-stranded RNAs targeting 109 genes expressed highly or specifically in mosquito hemocytes to identify novel regulators of the hemocyte immune response. Assays included phagocytosis of bacterial bioparticles, expression of the antimicrobial peptide CEC1, and basal and induced expression of the mosquito complement factor LRIM1. A cell viability screen was also carried out to assess dsRNA cytotoxicity and to identify genes involved in cell growth and survival. Our results identify 22 novel immune regulators, including proteins putatively involved in phagosome assembly and maturation (Ca2+ channel, v-ATPase and cyclin-dependent protein kinase), pattern recognition (fibrinogen-domain lectins and Nimrod), immune modulation (peptidase and serine protease homolog), immune signaling (Eiger and LPS-induced factor), cell adhesion and communication (Laminin B1 and Ninjurin) and immune homeostasis (Lipophorin receptor). The development of robust functional cell-based assays paves the way for genome-wide functional screens to study the mosquito immune response to infections with human pathogens. PMID:23382679

Anopheles gambiae feeding and survival on honeydew and extra-floral nectar of peridomestic plants.

PubMed

Gary, R E; Foster, W A

2004-06-01

It is widely believed that the malaria vector Anopheles gambiae Giles (Diptera: Culicidae) rarely or never feeds on sugar in nature. If so, the need for supplemental blood-feeding may be increased and this would help to explain why it is such an efficient malaria vector. Nonetheless, both sexes of this mosquito species readily imbibe and digest sugar solutions, and sugar is a staple of laboratory colonies. In this study, we investigated whether An. gambiae will feed on the extra-floral nectar of three common peridomestic plants in Africa, and on honeydew of the mealybug Pseudococcus longispinus (Targioni-Tozetti) (Hemiptera: Homoptera: Pseudococcidae), and how this affects survivorship. We found that both males and females of An. gambiae provided with vegetative parts of cassava (Manihot esculenta Crantz) survived as well (x = 26.3 and 19.2 days, respectively) as they did on 50% sucrose solution (x = 29.7 and 24.3 days, respectively) and much longer than they did on water alone (x = 1.8 days, both sexes). Females provided with mealybug honeydew also lived substantially longer (x = 16.5 days) than those on water alone. Males and females provided with vegetative parts of castorbean (Ricinus communis L.) also survived much longer (x = 12.7 and 7.8 days, respectively) than on water, but those provided with flowering lantana (Lantana camara L.) did not. Anthrone tests of females after one night of exposure to these potential energy sources confirmed that they obtained fructose from cassava, from mealybug honeydew, and from non-flowering castorbean, but not from lantana or from castorbean lacking its petiolar nectaries. Previous laboratory studies had shown that sugar availability affects the survival and biting frequency of An. gambiae. It now appears that this mosquito can locate natural sources of plant sugar readily and utilize them effectively. Nectar-producing plants in the domestic environment may play a significant role in this mosquito's energy budget and

Influence of biological and physicochemical characteristics of larval habitats on the body size of Anopheles gambiae mosquitoes (Diptera: Culicidae) along the Kenyan coast.

PubMed

Mwangangi, Joseph M; Mbogo, Charles M; Muturi, Ephantus J; Nzovua, Joseph G; Kabiru, Ephantus W; Githure, John I; Novak, Robert J; Beier, John C

2007-06-01

The number and productivity of larval habitats ultimately determine the density of adult mosquitoes. The biological and physicochemical conditions at the larval habitat affect larval development hence affecting the adult body size. The influence of biological and physicochemical characteristics on the body size of Anopheles gambiae was assessed in Jaribuni village, Kilifi district along the Kenyan Coast. Ten cages measuring 1 x 1 x 1 m (1 m3) with a netting material were placed in 10 different aquatic habitats, which were positive for anopheline mosquito larvae. Emergent mosquitoes were collected daily by aspiration and the wing lengths were determined by microscopy. In the habitats, physicochemical parameters were assessed: pH, surface debris, algae and emergent plants, turbidity, substrate, nitrate, ammonia, phosphate and chlorophyll a content. A total of 685 anopheline and culicine mosquitoes were collected from the emergent cages. Only female mosquitoes were considered in this study. Among the Anopheles spp, 202 were An. gambiae s.s., eight An. arabiensis, two An. funestus, whereas the Culex spp was composed of 214 Cx. quinquefasciatus, 10 Cx. tigripes, eight Cx. annulioris and one Cx. cumminsii. The mean wing length of the female An. gambiae s.s. mosquitoes was 3.02 mm (n=157), while that of An. arabiensis was 3.09 mm (n=9). There were no associations between the wing lengths and the environmental and chemical parameters, except for a positive correlation between wing length of An. gambiae and chlorophyll a content (r = 0.622). The day on which the mosquitoes emerged was not significant for the anopheline (p = 0.324) or culicine mosquitoes (p = 0.374), because the mosquito emerged from the cages on a daily basis. In conclusion, there was variability in production of emergent mosquitoes from different habitats, which means that there should be targeted control on these habitats based on productivity.

Efficacy of a novel mode of action of an indoor residual spraying product, SumiShield® 50WG against susceptible and resistant populations of Anopheles gambiae (s.l.) in Benin, West Africa.

PubMed

Agossa, Fiacre R; Padonou, Gil G; Koukpo, Come Z; Zola-Sahossi, Jacques; Azondekon, Roseric; Akuoko, Osei K; Ahoga, Juniace; N'dombidje, Boris; Akinro, Bruno; Fassinou, Arsene Jacques Y H; Sezonlin, Michel; Akogbeto, Martin C

2018-05-10

Scale-up of the distribution of long-lasting insecticide-treated bed nets and indoor residual spraying with insecticides over the last decade have contributed to the considerable decrease of malaria morbidity and mortality in sub-Saharan Africa. Due to the increasing pyrethroid resistance intensity and the spread of carbamate resistance in Anopheles gambiae (s.s.) mosquitoes and the limited number of insecticides recommended by the WHO for vector control, alternative insecticide formulations for IRS with long-lasting residual activity are required to sustain the gains obtained in most malaria-endemic countries. SumiShield 50WG (clothianidin 300 mg ai/m 2 ) developed by Sumitomo Chemical was evaluated alongside deltamethrin 25 mg ai/m 2 (K-Othrine 250 WG) against a pyrethroid resistant Anopheles gambiae (s.l.) population in experimental huts in Covè, Benin. Residual activity was also tested in cone bioassays with the susceptible An. gambiae "Kisumu" strain and the local wild resistant population. The results showed very low toxicity from deltamethrin (mortality rates ranged between 1-40%) against host-seeking resistant Anopheles populations. SumiShield in contrast gave an overall mean mortality of 91.7% at the 120 h observation across the eight- month observation period following spraying. The residual activity measured using cone tests was over the 80% WHO threshold for 24 weeks for resistant wild Anopheles population and 32 weeks for the susceptible strain "Kisumu" after the spraying. SumiShield is a good candidate for IRS in areas of permanent malaria transmission and where Anopheles populations are resistant to other conventional insecticides such as pyrethroids. It would be interesting to complete experimental huts studies by assessing the efficacy and residual effect of SumiShield 50WG at community level (small-scale field testing) in an area where vectors are highly resistant to insecticides.

Phylogenetic analysis of the GST family in Anopheles (Nyssorhynchus) darlingi.

PubMed

Azevedo-Júnior, Gilson Martins de; Guimarães-Marques, Giselle Moura; Cegatti Bridi, Leticia; Christine Ohse, Ketlen; Vicentini, Renato; Tadei, Wanderli; Rafael, Míriam Silva

2014-08-01

Anopheles darlingi Root, 1926 and Anopheles gambiae (Diptera: Culicidae) are the most important human malaria vectors in South America and Africa, respectively. The two species are estimated to have diverged 100 million years ago. Studies on the phylogenetics and evolution of gene sequences, such as glutathione S-transferase (GST) in disease-transmitting mosquitoes are scarce. The sigma class GST (KC890767) from the transcriptome of An. darlingi captured in the Brazilian Amazon was studied by in silico hybridization, and mapped to chromosome 3 of An. gambiae. The sigma class GST of An. darlingi was used for phylogenetic analyses to understand the GST base composition of the most recent common ancestor between An. darlingi, Anopheles gambiae, Aedes aegypti and Culex quinquefasciatus. The GST (KC890767) of An. darlingi was studied to generate the main divergence branches using a Neighbor-Joining and bootstrapping approaches to confirm confidence levels on the tree nodes that separate the An. darlingi and other mosquito species. The results showed divergence between An. gambiae, Ae. Aegypti, Cx. quinquefasciatus, and Phlebotomus papatasi as outgroup, and the homology relationship between sigma class GST of An. darlingi and GSTS1_1 gene of An. gambiae was valuable for phylogenetic and evolutionary studies. Copyright © 2014 Elsevier B.V. All rights reserved.

Select Small Core Structure Carbamates Exhibit High Contact Toxicity to “Carbamate-Resistant” Strain Malaria Mosquitoes, Anopheles gambiae (Akron)

PubMed Central

Wong, Dawn M.; Li, Jianyong; Chen, Qiao-Hong; Han, Qian; Mutunga, James M.; Wysinski, Ania; Anderson, Troy D.; Ding, Haizhen; Carpenetti, Tiffany L.; Verma, Astha; Islam, Rafique; Paulson, Sally L.; Lam, Polo C.-H.; Totrov, Maxim; Bloomquist, Jeffrey R.; Carlier, Paul R.

2012-01-01

Acetylcholinesterase (AChE) is a proven target for control of the malaria mosquito (Anopheles gambiae). Unfortunately, a single amino acid mutation (G119S) in An. gambiae AChE-1 (AgAChE) confers resistance to the AChE inhibitors currently approved by the World Health Organization for indoor residual spraying. In this report, we describe several carbamate inhibitors that potently inhibit G119S AgAChE and that are contact-toxic to carbamate-resistant An. gambiae. PCR-RFLP analysis was used to confirm that carbamate-susceptible G3 and carbamate-resistant Akron strains of An. gambiae carry wild-type (WT) and G119S AChE, respectively. G119S AgAChE was expressed and purified for the first time, and was shown to have only 3% of the turnover number (k cat) of the WT enzyme. Twelve carbamates were then assayed for inhibition of these enzymes. High resistance ratios (>2,500-fold) were observed for carbamates bearing a benzene ring core, consistent with the carbamate-resistant phenotype of the G119S enzyme. Interestingly, resistance ratios for two oxime methylcarbamates, and for five pyrazol-4-yl methylcarbamates were found to be much lower (4- to 65-fold). The toxicities of these carbamates to live G3 and Akron strain An. gambiae were determined. As expected from the enzyme resistance ratios, carbamates bearing a benzene ring core showed low toxicity to Akron strain An. gambiae (LC50>5,000 μg/mL). However, one oxime methylcarbamate (aldicarb) and five pyrazol-4-yl methylcarbamates (4a–e) showed good to excellent toxicity to the Akron strain (LC50 = 32–650 μg/mL). These results suggest that appropriately functionalized “small-core” carbamates could function as a resistance-breaking anticholinesterase insecticides against the malaria mosquito. PMID:23049714

Increased production of piRNAs from euchromatic clusters and genes in Anopheles gambiae compared with Drosophila melanogaster.

PubMed

George, Phillip; Jensen, Silke; Pogorelcnik, Romain; Lee, Jiyoung; Xing, Yi; Brasset, Emilie; Vaury, Chantal; Sharakhov, Igor V

2015-01-01

Specific genomic loci, termed Piwi-interacting RNA (piRNA) clusters, manufacture piRNAs that serve as guides for the inactivation of complementary transposable elements (TEs). The piRNA pathway has been accurately detailed in Drosophila melanogaster, while it remains poorly examined in other insects. This pathway is increasingly recognized as critical for germline development and reproduction. Understanding of the piRNA functions in mosquitoes could offer an opportunity for disease vector control by the reduction of their reproductive potential. To analyze the similarities and differences in this pathway between Drosophila and mosquito, we performed an in-depth analysis of the genomic loci producing piRNAs and their targets in the African malaria vector Anopheles gambiae. We identified 187 piRNA clusters in the An. gambiae genome and 155 piRNA clusters in the D. melanogaster genome. We demonstrate that many more piRNA clusters in the mosquito compared with the fruit fly are uni-directionally transcribed and are located outside pericentromeric heterochromatin. About 11 % of the An. gambiae piRNA population map to gene transcripts. This is a noticeable increase compared with the ~6 % of the piRNA population mapped to genes in D. melanogaster. A subset of the piRNA-enriched genes in An. gambiae has functions related to reproduction and development. At least 24 and 65 % of the mapped piRNAs correspond to genomic TE sequences in An. gambiae and D. melanogaster, respectively. DNA transposons and non-LTR retrotransposons are more abundant in An. gambiae, while LTR retrotransposons are more abundant in D. melanogaster. Yet, piRNAs predominantly target LTR retrotransposons in both species, which may point to a distinct feature of these elements compared to the other classes of TEs concerning their silencing by the piRNA pathway. Here, we demonstrate that piRNA-producing loci have more ubiquitous distribution in the An. gambiae genome than in the genome of D

Nigeria Anopheles Vector Database: An Overview of 100 Years' Research

PubMed Central

Okorie, Patricia Nkem; McKenzie, F. Ellis; Ademowo, Olusegun George; Bockarie, Moses; Kelly-Hope, Louise

2011-01-01

Anopheles mosquitoes are important vectors of malaria and lymphatic filariasis (LF), which are major public health diseases in Nigeria. Malaria is caused by infection with a protozoan parasite of the genus Plasmodium and LF by the parasitic worm Wuchereria bancrofti. Updating our knowledge of the Anopheles species is vital in planning and implementing evidence based vector control programs. To present a comprehensive report on the spatial distribution and composition of these vectors, all published data available were collated into a database. Details recorded for each source were the locality, latitude/longitude, time/period of study, species, abundance, sampling/collection methods, morphological and molecular species identification methods, insecticide resistance status, including evidence of the kdr allele, and P. falciparum sporozoite rate and W. bancrofti microfilaria prevalence. This collation resulted in a total of 110 publications, encompassing 484,747 Anopheles mosquitoes in 632 spatially unique descriptions at 142 georeferenced locations being identified across Nigeria from 1900 to 2010. Overall, the highest number of vector species reported included An. gambiae complex (65.2%), An. funestus complex (17.3%), An. gambiae s.s. (6.5%). An. arabiensis (5.0%) and An. funestus s.s. (2.5%), with the molecular forms An. gambiae M and S identified at 120 locations. A variety of sampling/collection and species identification methods were used with an increase in molecular techniques in recent decades. Insecticide resistance to pyrethroids and organochlorines was found in the main Anopheles species across 45 locations. Presence of P. falciparum and W. bancrofti varied between species with the highest sporozoite rates found in An. gambiae s.s, An. funestus s.s. and An. moucheti, and the highest microfilaria prevalence in An. gambiae s.l., An. arabiensis, and An. gambiae s.s. This comprehensive geo-referenced database provides an essential baseline on Anopheles

Genome-wide profiling of diel and circadian gene expression in the malaria vector Anopheles gambiae.

PubMed

Rund, Samuel S C; Hou, Tim Y; Ward, Sarah M; Collins, Frank H; Duffield, Giles E

2011-08-09

Anopheles gambiae, the primary African vector of malaria parasites, exhibits numerous rhythmic behaviors including flight activity, swarming, mating, host seeking, egg laying, and sugar feeding. However, little work has been performed to elucidate the molecular basis for these daily rhythms. To study how gene expression is regulated globally by diel and circadian mechanisms, we have undertaken a DNA microarray analysis of An. gambiae under light/dark cycle (LD) and constant dark (DD) conditions. Adult mated, non-blood-fed female mosquitoes were collected every 4 h for 48 h, and samples were processed with DNA microarrays. Using a cosine wave-fitting algorithm, we identified 1,293 and 600 rhythmic genes with a period length of 20-28 h in the head and body, respectively, under LD conditions, representing 9.7 and 4.5% of the An. gambiae gene set. A majority of these genes was specific to heads or bodies. Examination of mosquitoes under DD conditions revealed that rhythmic programming of the transcriptome is dependent on an interaction between the endogenous clock and extrinsic regulation by the LD cycle. A subset of genes, including the canonical clock components, was expressed rhythmically under both environmental conditions. A majority of genes had peak expression clustered around the day/night transitions, anticipating dawn and dusk. Genes cover diverse biological processes such as transcription/translation, metabolism, detoxification, olfaction, vision, cuticle regulation, and immunity, and include rate-limiting steps in the pathways. This study highlights the fundamental roles that both the circadian clock and light play in the physiology of this important insect vector and suggests targets for intervention.

Identification and validation of a gene causing cross-resistance between insecticide classes in Anopheles gambiae from Ghana.

PubMed

Mitchell, Sara N; Stevenson, Bradley J; Müller, Pie; Wilding, Craig S; Egyir-Yawson, Alexander; Field, Stuart G; Hemingway, Janet; Paine, Mark J I; Ranson, Hilary; Donnelly, Martin James

2012-04-17

In the last decade there have been marked reductions in malaria incidence in sub-Saharan Africa. Sustaining these reductions will rely upon insecticides to control the mosquito malaria vectors. We report that in the primary African malaria vector, Anopheles gambiae sensu stricto, a single enzyme, CYP6M2, confers resistance to two classes of insecticide. This is unique evidence in a disease vector of cross-resistance associated with a single metabolic gene that simultaneously reduces the efficacy of two of the four classes of insecticide routinely used for malaria control. The gene-expression profile of a highly DDT-resistant population of A. gambiae s.s. from Ghana was characterized using a unique whole-genome microarray. A number of genes were significantly overexpressed compared with two susceptible West African colonies, including genes from metabolic families previously linked to insecticide resistance. One of the most significantly overexpressed probe groups (false-discovery rate-adjusted P < 0.0001) belonged to the cytochrome P450 gene CYP6M2. This gene is associated with pyrethroid resistance in wild A. gambiae s.s. populations) and can metabolize both type I and type II pyrethroids in recombinant protein assays. Using in vitro assays we show that recombinant CYP6M2 is also capable of metabolizing the organochlorine insecticide DDT in the presence of solubilizing factor sodium cholate.

Identification and validation of a gene causing cross-resistance between insecticide classes in Anopheles gambiae from Ghana

PubMed Central

Mitchell, Sara N.; Stevenson, Bradley J.; Müller, Pie; Wilding, Craig S.; Egyir-Yawson, Alexander; Field, Stuart G.; Hemingway, Janet; Paine, Mark J. I.; Ranson, Hilary; Donnelly, Martin James

2012-01-01

In the last decade there have been marked reductions in malaria incidence in sub-Saharan Africa. Sustaining these reductions will rely upon insecticides to control the mosquito malaria vectors. We report that in the primary African malaria vector, Anopheles gambiae sensu stricto, a single enzyme, CYP6M2, confers resistance to two classes of insecticide. This is unique evidence in a disease vector of cross-resistance associated with a single metabolic gene that simultaneously reduces the efficacy of two of the four classes of insecticide routinely used for malaria control. The gene-expression profile of a highly DDT-resistant population of A. gambiae s.s. from Ghana was characterized using a unique whole-genome microarray. A number of genes were significantly overexpressed compared with two susceptible West African colonies, including genes from metabolic families previously linked to insecticide resistance. One of the most significantly overexpressed probe groups (false-discovery rate-adjusted P < 0.0001) belonged to the cytochrome P450 gene CYP6M2. This gene is associated with pyrethroid resistance in wild A. gambiae s.s. populations) and can metabolize both type I and type II pyrethroids in recombinant protein assays. Using in vitro assays we show that recombinant CYP6M2 is also capable of metabolizing the organochlorine insecticide DDT in the presence of solubilizing factor sodium cholate. PMID:22460795

Does the spillage of petroleum products in Anopheles breeding sites have an impact on the pyrethroid resistance?

PubMed Central

Djouaka, Rousseau F; Bakare, Adekunle A; Bankole, Honore S; Doannio, Julien MC; Coulibaly, Ousmane N; Kossou, Hortense; Tamo, Manuele; Basene, Harcourt I; Popoola, OK; Akogbeto, Martin C

2007-01-01

Background The emergence of Anopheles populations capable of withstanding lethal doses of insecticides has weakened the efficacy of most insecticide based strategies of vector control and, has highlighted the need for further studies on the mechanisms of insecticide resistance and the various factors selecting resistant populations of mosquitoes. This research targeted the analysis of breeding sites and the oviposition behaviour of susceptible and resistant populations of Anopheles in localities of spilled petroleum products. The aim was to establish the possible contribution of oil spillage in the selection of pyrethroid resistance in malaria vectors. Methods Anopheles breeding sites were identified and the insecticide susceptibility of the Anopheles gambiae populations mapped in 15 localities of South Western Nigeria. The presence of oil particles as well as the turbidity, the dissolved oxygen and the pH of each identified breeding site was recorded. Data were cross-analysed to correlate the habitat types and the insecticide susceptibility status of emerging mosquitoes. The second phase of this study was basically a laboratory model to provide more information on the implication of the spillage of petroleum on the selection of pyrethroid resistance in An. gambiae. Results Moderate levels of resistance following exposure to permethrin-impregnated papers were recorded with the majority of An. gambiae samples collected in the South Western Nigeria. Data from this study established a link between the constituency of the breeding sites and the resistance status of the emerging Anopheles. Conclusion This study has revealed the segregational occupation of breeding habitats by pyrethroid resistant and susceptible strains of An. gambiae in south-western Nigeria. Compiled results from field and laboratory research point out clear relationships between oil spillage and pyrethroid resistance in malaria vectors. The identification of this factor of resistance could serve as

No evidence for positive selection at two potential targets for malaria transmission-blocking vaccines in Anopheles gambiae s.s.

PubMed

Crawford, Jacob E; Rottschaefer, Susan M; Coulibaly, Boubacar; Sacko, Madjou; Niaré, Oumou; Riehle, Michelle M; Traore, Sékou F; Vernick, Kenneth D; Lazzaro, Brian P

2013-06-01

Human malaria causes nearly a million deaths in sub-Saharan Africa each year. The evolution of drug-resistance in the parasite and insecticide resistance in the mosquito vector has complicated control measures and made the need for new control strategies more urgent. Anopheles gambiae s.s. is one of the primary vectors of human malaria in Africa, and parasite-transmission-blocking vaccines targeting Anopheles proteins have been proposed as a possible strategy to control the spread of the disease. However, the success of these hypothetical technologies would depend on the successful ability to broadly target mosquito populations that may be genetically heterogeneous. Understanding the evolutionary pressures shaping genetic variation among candidate target molecules offers a first step towards evaluating the prospects of successfully deploying such technologies. We studied the population genetics of genes encoding two candidate target proteins, the salivary gland protein saglin and the basal lamina structural protein laminin, in wild populations of the M and S molecular forms of A. gambiae in Mali. Through analysis of intraspecific genetic variation and interspecific comparisons, we found no evidence of positive natural selection at the genes encoding these proteins. On the contrary, we found evidence for particularly strong purifying selection at the laminin gene. These results provide insight into the patterns of genetic diversity of saglin and laminin, and we discuss these findings in relation to the potential development of these molecules as vaccine targets. Copyright © 2013 Elsevier B.V. All rights reserved.

Comparative Genome and Proteome Analysis of Anopheles gambiae and Drosophila melanogaster

NASA Astrophysics Data System (ADS)

Zdobnov, Evgeny M.; von Mering, Christian; Letunic, Ivica; Torrents, David; Suyama, Mikita; Copley, Richard R.; Christophides, George K.; Thomasova, Dana; Holt, Robert A.; Subramanian, G. Mani; Mueller, Hans-Michael; Dimopoulos, George; Law, John H.; Wells, Michael A.; Birney, Ewan; Charlab, Rosane; Halpern, Aaron L.; Kokoza, Elena; Kraft, Cheryl L.; Lai, Zhongwu; Lewis, Suzanna; Louis, Christos; Barillas-Mury, Carolina; Nusskern, Deborah; Rubin, Gerald M.; Salzberg, Steven L.; Sutton, Granger G.; Topalis, Pantelis; Wides, Ron; Wincker, Patrick; Yandell, Mark; Collins, Frank H.; Ribeiro, Jose; Gelbart, William M.; Kafatos, Fotis C.; Bork, Peer

2002-10-01

Comparison of the genomes and proteomes of the two diptera Anopheles gambiae and Drosophila melanogaster, which diverged about 250 million years ago, reveals considerable similarities. However, numerous differences are also observed; some of these must reflect the selection and subsequent adaptation associated with different ecologies and life strategies. Almost half of the genes in both genomes are interpreted as orthologs and show an average sequence identity of about 56%, which is slightly lower than that observed between the orthologs of the pufferfish and human (diverged about 450 million years ago). This indicates that these two insects diverged considerably faster than vertebrates. Aligned sequences reveal that orthologous genes have retained only half of their intron/exon structure, indicating that intron gains or losses have occurred at a rate of about one per gene per 125 million years. Chromosomal arms exhibit significant remnants of homology between the two species, although only 34% of the genes colocalize in small ``microsyntenic'' clusters, and major interarm transfers as well as intra-arm shuffling of gene order are detected.

A spatial individual-based model predicting a great impact of copious sugar sources and resting sites on survival of Anopheles gambiae and malaria parasite transmission

USGS Publications Warehouse

Zhu, Lin; Qualls, Whitney A.; Marshall, John M; Arheart, Kris L.; DeAngelis, Donald L.; McManus, John W.; Traore, Sekou F.; Doumbia, Seydou; Schlein, Yosef; Muller, Gunter C.; Beier, John C.

2015-01-01

BackgroundAgent-based modelling (ABM) has been used to simulate mosquito life cycles and to evaluate vector control applications. However, most models lack sugar-feeding and resting behaviours or are based on mathematical equations lacking individual level randomness and spatial components of mosquito life. Here, a spatial individual-based model (IBM) incorporating sugar-feeding and resting behaviours of the malaria vector Anopheles gambiae was developed to estimate the impact of environmental sugar sources and resting sites on survival and biting behaviour.MethodsA spatial IBM containing An. gambiae mosquitoes and humans, as well as the village environment of houses, sugar sources, resting sites and larval habitat sites was developed. Anopheles gambiae behaviour rules were attributed at each step of the IBM: resting, host seeking, sugar feeding and breeding. Each step represented one second of time, and each simulation was set to run for 60 days and repeated 50 times. Scenarios of different densities and spatial distributions of sugar sources and outdoor resting sites were simulated and compared.ResultsWhen the number of natural sugar sources was increased from 0 to 100 while the number of resting sites was held constant, mean daily survival rate increased from 2.5% to 85.1% for males and from 2.5% to 94.5% for females, mean human biting rate increased from 0 to 0.94 bites per human per day, and mean daily abundance increased from 1 to 477 for males and from 1 to 1,428 for females. When the number of outdoor resting sites was increased from 0 to 50 while the number of sugar sources was held constant, mean daily survival rate increased from 77.3% to 84.3% for males and from 86.7% to 93.9% for females, mean human biting rate increased from 0 to 0.52 bites per human per day, and mean daily abundance increased from 62 to 349 for males and from 257 to 1120 for females. All increases were significant (P < 0.01). Survival was greater when sugar sources were randomly

Aquaporin water channel AgAQP1 in the malaria vector mosquito Anopheles gambiae during blood feeding and humidity adaptation

PubMed Central

Liu, Kun; Tsujimoto, Hitoshi; Cha, Sung-Jae; Agre, Peter; Rasgon, Jason L.

2011-01-01

Altered patterns of malaria endemicity reflect, in part, changes in feeding behavior and climate adaptation of mosquito vectors. Aquaporin (AQP) water channels are found throughout nature and confer high-capacity water flow through cell membranes. The genome of the major malaria vector mosquito Anopheles gambiae contains at least seven putative AQP sequences. Anticipating that transmembrane water movements are important during the life cycle of A. gambiae, we identified and characterized the A. gambiae aquaporin 1 (AgAQP1) protein that is homologous to AQPs known in humans, Drosophila, and sap-sucking insects. When expressed in Xenopus laevis oocytes, AgAQP1 transports water but not glycerol. Similar to mammalian AQPs, water permeation of AgAQP1 is inhibited by HgCl2 and tetraethylammonium, with Tyr185 conferring tetraethylammonium sensitivity. AgAQP1 is more highly expressed in adult female A. gambiae mosquitoes than in males. Expression is high in gut, ovaries, and Malpighian tubules where immunofluorescence microscopy reveals that AgAQP1 resides in stellate cells but not principal cells. AgAQP1 expression is up-regulated in fat body and ovary by blood feeding but not by sugar feeding, and it is reduced by exposure to a dehydrating environment (42% relative humidity). RNA interference reduces AgAQP1 mRNA and protein levels. In a desiccating environment (<20% relative humidity), mosquitoes with reduced AgAQP1 protein survive significantly longer than controls. These studies support a role for AgAQP1 in water homeostasis during blood feeding and humidity adaptation of A. gambiae, a major mosquito vector of human malaria in sub-Saharan Africa. PMID:21444767

Staggered larval time-to-hatch and insecticide resistance in the major malaria vector Anopheles gambiae S form.

PubMed

Kaiser, Maria L; Koekemoer, Lizette L; Coetzee, Maureen; Hunt, Richard H; Brooke, Basil D

2010-12-14

Anopheles gambiae is a major vector of malaria in the West African region. Resistance to multiple insecticides has been recorded in An. gambiae S form in the Ahafo region of Ghana. A laboratory population (GAH) established using wild material from this locality has enabled a mechanistic characterization of each resistance phenotype as well as an analysis of another adaptive characteristic - staggered larval time-to-hatch. Individual egg batches obtained from wild caught females collected from Ghana and the Republic of the Congo were monitored for staggered larval time-to-hatch. In addition, early and late larval time-to-hatch sub-colonies were selected from GAH. These selected sub-colonies were cross-mated and their hybrid progeny were subsequently intercrossed and back-crossed to the parental strains. The insecticide susceptibilities of the GAH base colony and the time-to-hatch selected sub-colonies were quantified for four insecticide classes using insecticide bioassays. Resistance phenotypes were mechanistically characterized using insecticide-synergist bioassays and diagnostic molecular assays for known reduced target-site sensitivity mutations. Anopheles gambiae GAH showed varying levels of resistance to all insecticide classes. Metabolic detoxification and reduced target-site sensitivity mechanisms were implicated. Most wild-caught families showed staggered larval time-to-hatch. However, some families were either exclusively early hatching or late hatching. Most GAH larvae hatched early but many egg batches contained a proportion of late hatching larvae. Crosses between the time-to-hatch selected sub-colonies yielded ambiguous results that did not fit any hypothetical models based on single-locus Mendelian inheritance. There was significant variation in the expression of insecticide resistance between the time-to-hatch phenotypes. An adaptive response to the presence of multiple insecticide classes necessarily involves the development of multiple resistance

Gravid Anopheles gambiae sensu stricto avoid ovipositing in Bermuda grass hay infusion and it's volatiles in two choice egg-count bioassays.

PubMed

Eneh, Lynda K; Okal, Michael N; Borg-Karlson, Anna-Karin; Fillinger, Ulrike; Lindh, Jenny M

2016-05-12

A number of mosquito species in the Culex and Aedes genera prefer to lay eggs in Bermuda grass (Cynodon dactylon) hay infusions compared to water alone. These mosquitoes are attracted to volatile compounds from the hay infusions making the infusions effective baits in gravid traps used for monitoring vectors of arboviral and filarial pathogens. Since Bermuda grass is abundant and widespread, it is plausible to explore infusions made from it as a potential low cost bait for outdoor monitoring of the elusive malaria vector Anopheles gambiae s.s. This study investigated preferential egg laying of individual An. gambiae s.s. in hay infusion or in tap water treated with volatiles detected in hay infusion headspace compared to tap water alone, using two-choice egg-count bioassays. Infusions were prepared by mixing 90 g of dried Bermuda grass (hay) with 24 L of unchlorinated tap water in a bucket, and leaving it for 3 days at ambient temperature and humidity. The volatiles in the headspace of the hay infusion were sampled with Tenax TA traps for 20 h and analysed using gas chromatography coupled to mass spectrometry. In total, 18 volatiles were detected in the infusion headspace. Nine of the detected compounds and nonanal were selected for bioassays. Eight of the selected compounds have previously been suggested to attract/stimulate egg laying in An. gambiae s.s. Gravid females were significantly (p < 0.05) less likely to lay eggs in hay infusion dilutions of 25, 50 and 100 % and in tap water containing any of six compounds (3-methylbutanol, phenol, 4-methylphenol, nonanal, indole, and 3-methylindole) compared to tap water alone. The oviposition response to 10 % hay infusion or any one of the remaining four volatiles (4-hepten-1-ol, phenylmethanol, 2-phenylethanol, or 4-ethylphenol) did not differ from that in tap water. Anopheles gambiae s.s. prefers to lay eggs in tap water rather than Bermuda grass hay infusion. This avoidance of the hay infusion appears to be

Dose–response tests and semi-field evaluation of lethal and sub-lethal effects of slow release pyriproxyfen granules (Sumilarv®0.5G) for the control of the malaria vectors Anopheles gambiae sensu lato

PubMed Central

2013-01-01

Background Recently research has shown that larviciding can be an effective tool for integrated malaria vector control. Nevertheless, the uptake of this intervention has been hampered by the need to re-apply larvicides frequently. There is a need to explore persistent, environmentally friendly larvicides for malaria vector control to reduce intervention efforts and costs by reducing the frequency of application. In this study, the efficacy of a 0.5% pyriproxyfen granule (Surmilarv®0.5G, Sumitomo Chemicals) was assessed for the control of Anopheles gambiae sensu stricto and Anopheles arabiensis, the major malaria vectors in sub-Saharan Africa. Methods Dose–response and standardized field tests were implemented following standard procedures of the World Health Organization’s Pesticide Evaluation Scheme to determine: (i) the susceptibility of vectors to this formulation; (ii) the residual activity and appropriate retreatment schedule for field application; and, (iii) sub-lethal impacts on the number and viability of eggs laid by adults after exposure to Sumilarv®0.5G during larval development. Results Anopheles gambiae s.s. and An. arabiensis were highly susceptible to Sumilarv®0.5G. Estimated emergence inhibition (EI) values were very low and similar for both species. The minimum dosage that completely inhibited adult emergence was between 0.01-0.03 parts per million (ppm) active ingredient (ai). Compared to the untreated control, an application of 0.018 ppm ai prevented 85% (95% confidence interval (CI) 82%-88%) of adult emergence over six weeks under standardized field conditions. A fivefold increase in dosage of 0.09 ppm ai prevented 97% (95% CI 94%-98%) emergence. Significant sub-lethal effects were observed in the standardized field tests. Female An. gambiae s.s. that were exposed to 0.018 ppm ai as larvae laid 47% less eggs, and females exposed to 0.09 ppm ai laid 74% less eggs than females that were unexposed to the treatment. Furthermore, 77

Spatial distribution and habitat characterisation of Anopheles larvae along the Kenyan coast

PubMed Central

Mwangangi, Joseph M.; Mbogo, Charles M.; Muturi, Ephantus J.; Nzovu, Joseph G.; Githure, John I.; Yan, Guiyun; Minakawa, Noboru; Novak, Robert; Beier, John C.

2009-01-01

Background & objectives A study was conducted to characterise larval habitats and to determine spatial heterogeneity of the Anopheles mosquito larvae. The study was conducted from May to June 1999 in nine villages along the Kenyan coast. Methods Aquatic habitats were sampled by use of standard dipping technique. The habitats were characterised based on size, pH, distance to the nearest house, coverage of canopy, surface debris, algae and emergent plants, turbidity, substrate, and habitat type. Results A total of 110 aquatic habitats like stream pools (n = 10); puddles (n = 65); tire tracks (n = 5); ponds (n = 5) and swamps (n = 25) were sampled in nine villages located in three districts of the Kenyan coast. A total of 7,263 Anopheles mosquito larvae were collected, 63.9% were early instars and 36.1% were late instars. Morphological identification of the III and IV instar larvae by use of microscopy yielded 90.66% (n = 2,377) Anopheles gambiae Complex, 0.88% (n = 23) An. funestus, An. coustani 7.63% (n = 200), An. rivulorum 0.42% (n = 11), An. pharoensis 0.19% (n = 5), An. swahilicus 0.08% (n = 2), An. wilsoni 0.04% (n = 1) and 0.11% (n = 3) were unidentified. A subset of the An. gambiae Complex larvae identified morphologically, was further analysed using rDNA-PCR technique resulting in 68.22% (n = 1,290) An. gambiae s.s., 7.93% (n = 150) An. arabiensis and 23.85% (n = 451) An. merus. Multiple logistic regression model showed that emergent plants (p = 0.019), and floating debris (p = 0.038) were the best predictors of An. gambiae larval abundance in these habitats. Interpretation & conclusion Habitat type, floating debris and emergent plants were found to be the key factors determining the presence of Anopheles larvae in the habitats. For effective larval control, the type of habitat should be considered and most productive habitat type be given a priority in the mosquito abatement programme. PMID:17378216

Genetic Dissection of Anopheles gambiae Gut Epithelial Responses to Serratia marcescens

PubMed Central

Stathopoulos, Stavros; Neafsey, Daniel E.; Lawniczak, Mara K. N.; Muskavitch, Marc A. T.; Christophides, George K.

2014-01-01

Genetic variation in the mosquito Anopheles gambiae profoundly influences its ability to transmit malaria. Mosquito gut bacteria are shown to influence the outcome of infections with Plasmodium parasites and are also thought to exert a strong drive on genetic variation through natural selection; however, a link between antibacterial effects and genetic variation is yet to emerge. Here, we combined SNP genotyping and expression profiling with phenotypic analyses of candidate genes by RNAi-mediated silencing and 454 pyrosequencing to investigate this intricate biological system. We identified 138 An. gambiae genes to be genetically associated with the outcome of Serratia marcescens infection, including the peptidoglycan recognition receptor PGRPLC that triggers activation of the antibacterial IMD/REL2 pathway and the epidermal growth factor receptor EGFR. Silencing of three genes encoding type III fibronectin domain proteins (FN3Ds) increased the Serratia load and altered the gut microbiota composition in favor of Enterobacteriaceae. These data suggest that natural genetic variation in immune-related genes can shape the bacterial population structure of the mosquito gut with high specificity. Importantly, FN3D2 encodes a homolog of the hypervariable pattern recognition receptor Dscam, suggesting that pathogen-specific recognition may involve a broader family of immune factors. Additionally, we showed that silencing the gene encoding the gustatory receptor Gr9 that is also associated with the Serratia infection phenotype drastically increased Serratia levels. The Gr9 antibacterial activity appears to be related to mosquito feeding behavior and to mostly rely on changes of neuropeptide F expression, together suggesting a behavioral immune response following Serratia infection. Our findings reveal that the mosquito response to oral Serratia infection comprises both an epithelial and a behavioral immune component. PMID:24603764

Repellent, irritant and toxic effects of 20 plant extracts on adults of the malaria vector Anopheles gambiae mosquito.

PubMed

Deletre, Emilie; Martin, Thibaud; Campagne, Pascal; Bourguet, Denis; Cadin, Andy; Menut, Chantal; Bonafos, Romain; Chandre, Fabrice

2013-01-01

Pyrethroid insecticides induce an excito-repellent effect that reduces contact between humans and mosquitoes. Insecticide use is expected to lower the risk of pathogen transmission, particularly when impregnated on long-lasting treated bednets. When applied at low doses, pyrethroids have a toxic effect, however the development of pyrethroid resistance in several mosquito species may jeopardize these beneficial effects. The need to find additional compounds, either to kill disease-carrying mosquitoes or to prevent mosquito contact with humans, therefore arises. In laboratory conditions, the effects (i.e., repellent, irritant and toxic) of 20 plant extracts, mainly essential oils, were assessed on adults of Anopheles gambiae, a primary vector of malaria. Their effects were compared to those of DEET and permethrin, used as positive controls. Most plant extracts had irritant, repellent and/or toxic effects on An. gambiae adults. The most promising extracts, i.e. those combining the three types of effects, were from Cymbopogon winterianus, Cinnamomum zeylanicum and Thymus vulgaris. The irritant, repellent and toxic effects occurred apparently independently of each other, and the behavioural response of adult An. gambiae was significantly influenced by the concentration of the plant extracts. Mechanisms underlying repellency might, therefore, differ from those underlying irritancy and toxicity. The utility of the efficient plant extracts for vector control as an alternative to pyrethroids may thus be envisaged.

Repellent, Irritant and Toxic Effects of 20 Plant Extracts on Adults of the Malaria Vector Anopheles gambiae Mosquito

PubMed Central

Deletre, Emilie; Martin, Thibaud; Campagne, Pascal; Bourguet, Denis; Cadin, Andy; Menut, Chantal; Bonafos, Romain; Chandre, Fabrice

2013-01-01

Pyrethroid insecticides induce an excito-repellent effect that reduces contact between humans and mosquitoes. Insecticide use is expected to lower the risk of pathogen transmission, particularly when impregnated on long-lasting treated bednets. When applied at low doses, pyrethroids have a toxic effect, however the development of pyrethroid resistance in several mosquito species may jeopardize these beneficial effects. The need to find additional compounds, either to kill disease-carrying mosquitoes or to prevent mosquito contact with humans, therefore arises. In laboratory conditions, the effects (i.e., repellent, irritant and toxic) of 20 plant extracts, mainly essential oils, were assessed on adults of Anopheles gambiae, a primary vector of malaria. Their effects were compared to those of DEET and permethrin, used as positive controls. Most plant extracts had irritant, repellent and/or toxic effects on An. gambiae adults. The most promising extracts, i.e. those combining the three types of effects, were from Cymbopogon winterianus, Cinnamomum zeylanicum and Thymus vulgaris. The irritant, repellent and toxic effects occurred apparently independently of each other, and the behavioural response of adult An. gambiae was significantly influenced by the concentration of the plant extracts. Mechanisms underlying repellency might, therefore, differ from those underlying irritancy and toxicity. The utility of the efficient plant extracts for vector control as an alternative to pyrethroids may thus be envisaged. PMID:24376515

Comparison of male reproductive success in malaria-refractory and susceptible strains of Anopheles gambiae

PubMed Central

Voordouw, Maarten J; Koella, Jacob C; Hurd, Hilary

2008-01-01

Background In female mosquitoes that transmit malaria, the benefits of being refractory to the Plasmodium parasite are balanced by the immunity costs in the absence of infection. Male mosquitoes, however, gain no advantage from being refractory to blood-transmitted parasites, so that any costs associated with an enhanced immune system in the males limit the evolution of female refractoriness and has practical implications for the release of transgenic males. Methods Aspects of the male cost of carrying Plasmodium-refractory genes were estimated by comparing the males' immune response and reproductive success among strains of Anopheles gambiae that had been selected for refractoriness or extreme susceptibility to the rodent malaria parasite, Plasmodium yoelii nigeriensis. The refractory males had a stronger melanization response than males from the susceptible line. Four traits were used as correlates of a male's reproductive success: the proportion of females that were inseminated by a fixed number of males in a cage within a fixed time frame, the proportion of females with motile sperm in their spermathecae, the proportion of ovipositing females, and the mean number of eggs per batch. Results Although there were significant differences among groups of males in sperm motility and oviposition success, these differences in male reproductive success were not associated with the refractory or susceptible male genotypes. Contrary to expectation, females mated to early emerging refractory males laid significantly more eggs per batch than females mated to later emerging susceptible males. Sperm motility and oviposition success were strongly correlated suggesting that variation in sperm motility influences female oviposition and ultimately male reproductive success. Conclusion An increased melanization response in male A. gambiae does not diminish male reproductive success under the experimental protocol used in this study. That refractory males induced ovipositing females

Dramatic Decrease in Malaria Transmission after Large-Scale Indoor Residual Spraying with Bendiocarb in Benin, an Area of High Resistance of Anopheles gambiae to Pyrethroids

PubMed Central

Akogbeto, Martin; Padonou, Gil Germain; Bankole, Honore Sourou; Gazard, Dorothee Kinde; Gbedjissi, Ghelus Louis

2011-01-01

In 2008, the National Malaria Control Program in Benin implemented a vector control intervention based on indoor residual spraying (IRS). Four districts of high resistance of Anopheles gambiae to pyrethroids were sprayed with bendiocarb. More than 350,000 inhabitants have been protected. Entomologic parameters in the control area were compared with those in intervention sites. The study has shown a drastic decrease in the An. gambiae biting rate in the sprayed areas. Results of an enzyme-linked immunosorbent assay were negative for Plasmodium falciparum antigen during the entire period of the intervention. No household members received infected bites (entomologic inoculation rate = 0 during January–July). Parous rates were low in areas covered by IRS because bendiocarb is not conducive to long-term mosquito survival. Bendiocarb was found to be a good alternative insecticide for IRS in Benin, in areas where An. gambiae has developed high resistance to pyrethroids. PMID:21976555

New evidence of mating swarms of the malaria vector, Anopheles arabiensis in Tanzania

PubMed Central

Kaindoa, Emmanuel W.; Ngowo, Halfan S.; Limwagu, Alex; Mkandawile, Gustav; Kihonda, Japhet; Masalu, John Paliga; Bwanary, Hamis; Diabate, Abdoulaye; Okumu, Fredros O.

2017-01-01

Background: Malaria mosquitoes form mating swarms around sunset, often at the same locations for months or years. Unfortunately, studies of Anopheles swarms are rare in East Africa, the last recorded field observations in Tanzania having been in 1983. Methods: Mosquito swarms were surveyed by trained volunteers between August-2016 and June-2017 in Ulanga district, Tanzania. Identified Anopheles swarms were sampled using sweep nets, and collected mosquitoes killed by refrigeration then identified by sex and taxa. Sub-samples were further identified by PCR, and spermatheca of females examined for mating status. Mosquito ages were estimated by observing female ovarian tracheoles and rotation of male genitalia. GPS locations, types of swarm markers, start/end times of swarming, heights above ground, mosquito counts/swarm, and copulation events were recorded. Results: A total of 216 Anopheles swarms were identified, characterized and mapped, from which 7,142 Anopheles gambiae s.l and 13 Anopheles funestus were sampled. The An. gambiae s.l were 99.6% males and 0.4% females, while the An. funestus were all males. Of all An. gambiae s.l analyzed by PCR, 86.7% were An. arabiensis, while 13.3% returned non-amplified DNA. Mean height (±SD) of swarms was 2.74±0.64m, and median duration was 20 (IQR; 15-25) minutes. Confirmed swarm markers included rice fields (25.5%), burned grounds (17.2%), banana trees (13%), brick piles (8.8%), garbage heaps (7.9%) and ant-hills (7.4%). Visual estimates of swarm sizes by the volunteers was strongly correlated to actual sizes by sweep nets (R=0.94; P=<0.001). All females examined were nulliparous and 95.6% [N=6787] of males had rotated genitalia, indicating sexual maturity. Conclusions: This is the first report of Anopheles swarms in Tanzania in more than three decades. The study demonstrates that the swarms can be identified and characterized by trained community-based volunteers, and highlights potential new interventions, for example

The relationship between Anopheles gambiae density and rice cultivation in the savannah zone and forest zone of Côte d'Ivoire.

PubMed

Briët, Olivier J T; Dossou-Yovo, Joel; Akodo, Elena; van de Giesen, Nick; Teuscher, Thomas M

2003-05-01

In 13 villages in the savannah zone and 21 villages in the forest zone of Côte d'Ivoire, the biting density of the principal malaria vector, Anopheles gambiae, was studied as a function of rice cultivation in the inland valleys in a 2-km radius around each village. In the savannah villages, during the main season cropping period, surface water on rice-cultivated and to a lesser extent on uncultivated inland valleys seems to contribute strongly to the A. gambiae population density. For the off-season cropping period (which starts after the first light rains in the savannah zone), correlations were weaker. Breeding sites other than in inland valleys may play an important role in the savannah zone. In the forest zone, however, the A. gambiae population density was strongly correlated with the surface water availability (SWA) in the rice-cultivated inland valleys, whereas the correlation with the SWA in other (uncultivated) inland valleys was weak. The requirement of sunlit breeding sites for A. gambiae might explain this difference between zones. In the forest zone, only inland valleys cleared for rice cultivation meet this requirement, whereas all other inland valleys are covered with dense vegetation. In the savannah zone, however, most undergrowth is burnt during the dry season, which permits sunlight to reach puddles resulting from the first rains.

Incorporating the effects of humidity in a mechanistic model of Anopheles gambiae mosquito population dynamics in the Sahel region of Africa

PubMed Central

2013-01-01

Background Low levels of relative humidity are known to decrease the lifespan of mosquitoes. However, most current models of malaria transmission do not account for the effects of relative humidity on mosquito survival. In the Sahel, where relative humidity drops to levels <20% for several months of the year, we expect relative humidity to play a significant role in shaping the seasonal profile of mosquito populations. Here, we present a new formulation for Anopheles gambiae sensu lato (s.l.) mosquito survival as a function of temperature and relative humidity and investigate the effect of humidity on simulated mosquito populations. Methods Using existing observations on relationships between temperature, relative humidity and mosquito longevity, we developed a new equation for mosquito survival as a function of temperature and relative humidity. We collected simultaneous field observations on temperature, wind, relative humidity, and anopheline mosquito populations for two villages from the Sahel region of Africa, which are presented in this paper. We apply this equation to the environmental data and conduct numerical simulations of mosquito populations using the Hydrology, Entomology and Malaria Transmission Simulator (HYDREMATS). Results Relative humidity drops to levels that are uncomfortable for mosquitoes at the end of the rainy season. In one village, Banizoumbou, water pools dried up and interrupted mosquito breeding shortly after the end of the rainy season. In this case, relative humidity had little effect on the mosquito population. However, in the other village, Zindarou, the relatively shallow water table led to water pools that persisted several months beyond the end of the rainy season. In this case, the decrease in mosquito survival due to relative humidity improved the model’s ability to reproduce the seasonal pattern of observed mosquito abundance. Conclusions We proposed a new equation to describe Anopheles gambiae s.l. mosquito survival as a

Successful field trial of attractive toxic sugar bait (ATSB) plant-spraying methods against malaria vectors in the Anopheles gambiae complex in Mali, West Africa

PubMed Central

2010-01-01

Background Based on highly successful demonstrations in Israel that attractive toxic sugar bait (ATSB) methods can decimate local populations of mosquitoes, this study determined the effectiveness of ATSB methods for malaria vector control in the semi-arid Bandiagara District of Mali, West Africa. Methods Control and treatment sites, selected along a road that connects villages, contained man-made ponds that were the primary larval habitats of Anopheles gambiae and Anopheles arabiensis. Guava and honey melons, two local fruits shown to be attractive to An. gambiae s.l., were used to prepare solutions of Attractive Sugar Bait (ASB) and ATSB that additionally contained boric acid as an oral insecticide. Both included a color dye marker to facilitate determination of mosquitoes feeding on the solutions. The trial was conducted over a 38-day period, using CDC light traps to monitor mosquito populations. On day 8, ASB solution in the control site and ATSB solution in the treatment site were sprayed using a hand-pump on patches of vegetation. Samples of female mosquitoes were age-graded to determine the impact of ATSB treatment on vector longevity. Results Immediately after spraying ATSB in the treatment site, the relative abundance of female and male An. gambiae s.l. declined about 90% from pre-treatment levels and remained low. In the treatment site, most females remaining after ATSB treatment had not completed a single gonotrophic cycle, and only 6% had completed three or more gonotrophic cycles compared with 37% pre-treatment. In the control site sprayed with ASB (without toxin), the proportion of females completing three or more gonotrophic cycles increased from 28.5% pre-treatment to 47.5% post-treatment. In the control site, detection of dye marker in over half of the females and males provided direct evidence that the mosquitoes were feeding on the sprayed solutions. Conclusion This study in Mali shows that even a single application of ATSB can substantially

Independent and interactive effect of plant- and mammalian- based odors on the response of the malaria vector, Anopheles gambiae.

PubMed

Jacob, Juliah W; Tchouassi, David P; Lagat, Zipporah O; Mathenge, Evan M; Mweresa, Collins K; Torto, Baldwyn

2018-04-27

Several studies have shown that odors of plant and animal origin can be developed into lures for use in surveillance of mosquito vectors of infectious diseases. However, the effect of combining plant- and mammalian-derived odors into an improved lure for monitoring both nectar- and blood-seeking mosquito populations in traps is yet to be explored. Here we used both laboratory dual choice olfactometer and field assays to investigate responses of the malaria vector, Anopheles gambiae, to plant- and mammalian-derived compounds and a combined blend derived from these two odor sources. Using subtractive bioassays in dual choice olfactometer we show that a 3-component terpenoid plant-derived blend comprising (E)-linalool oxide, β-pinene, β-ocimene was more attractive to females of An. gambiae than (E)-linalool oxide only (previously found attractive in field trials) and addition of limonene to this blend antagonized its attractiveness. Likewise, a mammalian-derived lure comprising the aldehydes heptanal, octanal, nonanal and decanal, was more preferred than (E)-linalool oxide. Surprisingly, combining the plant-derived 3-component blend with the mammalian derived 4-component blend attracted fewer females of An. gambiae than the individual blends in laboratory assays. However, this pattern was not replicated in field trials, where we observed a dose-dependent effect on trap catches while combining both blends with significantly improved trap catches at higher doses. The observed dose-dependent attractiveness for An. gambiae has practical implication in the design of vector control strategies involving kairomones from plant- and mammalian-based sources. Copyright © 2018 Elsevier B.V. All rights reserved.

Involvement of the Anopheles gambiae Nimrod gene family in mosquito immune responses.

PubMed

Estévez-Lao, Tania Y; Hillyer, Julián F

2014-01-01

Insects fight infection using a variety of signaling pathways and immune effector proteins. In Drosophila melanogaster, three members of the Nimrod gene family (draper, nimC1 and eater) bind bacteria, and this binding leads to phagocytosis by hemocytes. The Nimrod gene family has since been identified in other insects, but their function in non-drosophilids remains unknown. The purpose of this study was to identify the members of the Nimrod gene family in the malaria mosquito, Anopheles gambiae, and to assess their role in immunity. We identified and sequenced three members of this gene family, herein named draper, nimrod and eater, which are the orthologs of D. melanogaster draper, nimB2 and eater, respectively. The three genes are preferentially expressed in hemocytes and their peak developmental expression is in pupae and young adults. Infection induces the transcriptional upregulation of all three genes, but the magnitude of this upregulation becomes more attenuated as mosquitoes become older. RNAi-based knockdown of eater, but not draper or nimrod, decreased a mosquito's ability to kill Escherichia coli in the hemocoel. Knockdown of draper, eater, or any combination of Nimrod family genes rendered mosquitoes more likely to die from Staphylococcus epidermidis. Finally, knockdown of Nimrod family genes did not impact mRNA levels of the antimicrobial peptides defensin (def1), cecropin (cecA) or gambicin (gam1), but eater knockdown led to a decrease in mRNA levels of nitric oxide synthase. Together, these data show that members of the A. gambiae Nimrod gene family are positive regulators of the mosquito antibacterial response. Copyright © 2013 Elsevier Ltd. All rights reserved.

Hemolymph proteins of Anopheles gambiae larvae infected by Escherichia coli.

PubMed

He, Xuesong; Cao, Xiaolong; He, Yan; Bhattarai, Krishna; Rogers, Janet; Hartson, Steve; Jiang, Haobo

2017-09-01

Anopheles gambiae is a major vector of human malaria and its immune system in part determines the fate of ingested parasites. Proteins, hemocytes and fat body in hemolymph are critical components of this system, mediating both humoral and cellular defenses. Here we assessed differences in the hemolymph proteomes of water- and E. coli-pricked mosquito larvae by a gel-LC-MS approach. Among the 1756 proteins identified, 603 contained a signal peptide but accounted for two-third of the total protein amount on the quantitative basis. The sequence homology search indicated that 233 of the 1756 may be related to defense. In general, we did not detect substantial differences between the control and induced plasma samples in terms of protein numbers or levels. Protein distributions in the gel slices suggested post-translational modifications (e.g. proteolysis) and formation of serpin-protease complexes and high Mr immune complexes. Based on the twenty-five most abundant proteins, we further suggest that major functions of the larval hemolymph are storage, transport, and immunity. In summary, this study provided first data on constitution, levels, and possible functions of hemolymph proteins in the mosquito larvae, reflecting complex changes occurring in the fight against E. coli infection. Copyright © 2017 Elsevier Ltd. All rights reserved.

Cotton pest management practices and the selection of pyrethroid resistance in Anopheles gambiae population in northern Benin.

PubMed

Yadouleton, Anges; Martin, Thibaud; Padonou, Gil; Chandre, Fabrice; Asidi, Alex; Djogbenou, Luc; Dabiré, Roch; Aïkpon, Rock; Boko, Michel; Glitho, Isabelle; Akogbeto, Martin

2011-04-13

Pyrethroid insecticides, carbamate and organophosphate are the classes of insecticides commonly used in agriculture for crop protection in Benin. Pyrethroids remain the only class of insecticides recommended by the WHO for impregnation of bed nets. Unfortunately, the high level of pyrethroid resistance in Anopheles gambiae s.l., threatens to undermine the success of pyrethroid treated nets. This study focuses on the investigation of agricultural practices in cotton growing areas, and their direct impact on larval populations of An. gambiae in surrounding breeding sites. The protocol was based on the collection of agro-sociological data where farmers were subjected to semi-structured questionnaires based on the strategies used for crop protection. This was complemented by bioassay tests to assess the susceptibility of malaria vectors to various insecticides. Molecular analysis was performed to characterize the resistance genes and the molecular forms of An. gambiae. Insecticide residues in soil samples from breeding sites were investigated to determine major factors that can inhibit the normal growth of mosquito larvae by exposing susceptible and resistant laboratory strains. There is a common use by local farmers of mineral fertilizer NPK at 200 kg/ha and urea at 50 kg/hectare following insecticide treatments in both the Calendar Control Program (CCP) and the Targeted Intermittent Control Program (TICP). By contrast, no chemicals are involved in Biological Program (BP) where farmers use organic and natural fertilizers which include animal excreta.Susceptibility test results confirmed a high resistance to DDT. Mean mortality of An. gambiae collected from the farms practicing CCP, TICP and BP methods were 33%, 42% and 65% respectively. An. gambiae populations from areas using the CCP and TICP programs showed resistance to permethrin with mortality of 50% and 58% respectively. By contrast, bioassay test results of An. gambiae from BP areas gave a high level of

Evidence of multiple insecticide resistance mechanisms in Anopheles gambiae populations in Bangui, Central African Republic.

PubMed

Olé Sangba, Marina Lidwine; Sidick, Aboubakar; Govoetchan, Renaud; Dide-Agossou, Christian; Ossè, Razaki A; Akogbeto, Martin; Ndiath, Mamadou Ousmane

2017-01-13

Knowledge of insecticide resistance status in the main malaria vectors is an essential component of effective malaria vector control. This study presents the first evaluation of the status of insecticide resistance in Anopheles gambiae populations from Bangui, the Central African Republic. Anopheles mosquitoes were reared from larvae collected in seven districts of Bangui between September to November 2014. The World Health Organisation's bioassay susceptibility tests to lambda-cyhalothrin (0.05%), deltamethrin (0.05%), DDT (4%), malathion (5%), fenitrothion (1%) and bendiocarb (0.1%) were performed on adult females. Species and molecular forms as well as the presence of L1014F kdr and Ace-1 R mutations were assessed by PCR. Additional tests were conducted to assess metabolic resistance status. After 1 h exposure, a significant difference of knockdown effect was observed between districts in all insecticides tested except deltamethrin and malathion. The mortality rate (MR) of pyrethroids group ranging from 27% (CI: 19-37.5) in Petevo to 86% (CI: 77.6-92.1) in Gbanikola; while for DDT, MR ranged from 5% (CI: 1.6-11.3) in Centre-ville to 39% (CI: 29.4-49.3) in Ouango. For the organophosphate group a MR of 100% was observed in all districts except Gbanikola where a MR of 96% (CI: 90-98.9) was recorded. The mortality induced by bendiocarb was very heterogeneous, ranging from 75% (CI: 62.8-82.8) in Yapele to 99% (CI: 84.5-100) in Centre-ville. A high level of kdr-w (L1014F) frequency was observed in all districts ranging from 93 to 100%; however, no kdr-e (L1014S) and Ace-1 R mutation were found in all tested mosquitoes. Data of biochemical analysis showed significant overexpression activities of cytochrome P450, GST and esterases in Gbanikola and Yapele (χ 2  = 31.85, df = 2, P < 0.001). By contrast, esterases activities using α and β-naphthyl acetate were significantly low in mosquitoes from PK10 and Ouango in comparison to Kisumu strain (χ 2 �

Evaluation of the influence of electric nets on the behaviour of oviposition site seeking Anopheles gambiae s.s

PubMed Central

2014-01-01

Background Electric nets (e-nets) are used to analyse the flight behaviour of insects and have been used extensively to study the host-oriented flight of tsetse flies. Recently we adapted this tool to analyse the oviposition behaviour of gravid malaria vectors, Anopheles gambiae s.s., orienting towards aquatic habitats and traps by surrounding an artificial pond with e-nets and collecting electrocuted mosquitoes on sticky boards on the ground next to the nets. Here we study whether e-nets themselves affect the responses of gravid An. gambiae s.s.. Methods Dual-choice experiments were carried out in 80 m2 screened semi-field systems where 200 gravid An. gambiae s.s. were released each night for 12 nights per experiment. The numbers of mosquito landing on or approaching an oviposition site were studied by adding detergent to the water in an artificial pond or surrounding the pond with a square of e-nets. We also assessed whether the supporting framework of the nets or the sticky boards used to retain electrocuted mosquitoes influenced the catch. Results Two similar detergent treated ponds presented in choice tests caught an equal proportion of the mosquitoes released, whereas a pond surrounded by e-nets caught a higher proportion than an open pond (odds ratio (OR) 1.7, 95% confidence interval (CI) 1.1 - 2.7; p < 0.017). The separate evaluation of the impact of the square of electric nets and the yellow boards on the approach of gravid females towards a pond suggests that the tower-like construction of the square of electric nets did not restrict the approach of females but the yellow sticky boards on the ground attract gravid females to a source of water (OR 2.7 95% CI 1.7 – 4.3; p < 0.001). Conclusion The trapping efficiency of the electric nets is increased when large yellow sticky boards are placed on the ground next to the e-nets to collect electrocuted mosquitoes, possibly because of increased visual contrast to the aquatic habitat. It is therefore

Functional characterisation of the Anopheles leucokinins and their cognate G-protein coupled receptor.

PubMed

Radford, Jonathan C; Terhzaz, Selim; Cabrero, Pablo; Davies, Shireen-A; Dow, Julian A T

2004-12-01

Identification of the Anopheles gambiae leucokinin gene from the completed A. gambiae genome revealed that this insect species contains three leucokinin peptides, named Anopheles leucokinin I-III. These peptides are similar to those identified in two other mosquito species, Aedes aegypti and Culex salinarius. Additionally, Anopheles leucokinin I displays sequence similarity to Drosophila melanogaster leucokinin. Using a combination of computational and molecular approaches, a full-length cDNA for a candidate leucokinin-like receptor was isolated from A. stephensi, a close relative of A. gambiae. Alignment of the known leucokinin receptors--all G protein-coupled receptors (GPCRs)--with this receptor, identified some key conserved regions within the receptors, notably transmembrane (TM) domains I, II, III, VI and VII. The Anopheles leucokinins and receptor were shown to be a functional receptor-ligand pair. All three Anopheles leucokinins caused a dose-dependent rise in intracellular calcium ([Ca2+]i) when applied to S2 cells co-expressing the receptor and an aequorin transgene, with a potency order of I>II>III. Drosophila leucokinin was also found to activate the Anopheles receptor with a similar EC50 value to Anopheles leucokinin I. However, when the Anopheles peptides were applied to the Drosophila receptor, only Anopheles leucokinin I and II elicited a rise in [Ca2+]i. This suggests that the Anopheles receptor has a broader specificity for leucokinin ligands than the Drosophila receptor. Antisera raised against the Anopheles receptor identified a doublet of approx. 65 and 72 kDa on western blots, consistent with the presence of four N-glycosylation sites within the receptor sequence, and the known glycosylation of the receptor in Drosophila. In Anopheles tubules, as in Drosophila, the receptor was localised to the stellate cells. Thus we provide the first identification of Anopheles mosquito leucokinins (Anopheles leucokinins) and a cognate leucokinin receptor

Constituents of the Essential Oil of Suregada zanzibariensis Leaves are Repellent to the Mosquito, Anopheles gambiae s.s.

PubMed Central

Innocent, Ester; Joseph, Cosam C.; Gikonyo, Nicholas K.; Nkunya, Mayunga H.H.; Hassanali, Ahmed

2010-01-01

In traditional African communities, repellent volatiles from certain plants generated by direct burning or by thermal expulsion have played an important role in protecting households against vectors of malaria and other diseases. Previous research on volatile constituents of plants has shown that some are good sources of potent mosquito repellents. In this bioprospecting initiative, the essential oil of leaves of the tree, Suregada zanzibariensis Verdc. (Angiospermae: Euphobiaceae) was tested against the mosquito, Anopheles gambiae s.s. Giles (Diptera: Culicidae) and found to be repellent. Gas chromatography (GC), GC-linked mass spectrometry (GC-MS) and, where possible, GC-co-injections with authentic compounds, led to the identification of about 34 compounds in the essential oil. About 56% of the constituents were terpenoid ketones, mostly methyl ketones. Phenylacetaldehyde (14.4%), artemisia ketone (10.1%), (1S)-(-)-verbenone (12.1%) and geranyl acetone (9.4%) were the main constituents. Apart from phenylacetaldehyde, repellent activities of the other main constituents were higher than that of the essential oil. The blends of the main constituents in proportions found in the essential oil were more repellent to An. gambiae s.s. than was the parent oil (p < 0.05), and the presence of artemisia ketone in the blend caused a significant increase in the repellency of the resulting blend. These results suggested that blends of some terpenoid ketones can serve as effective An. gambiae s.s. mosquito repellents. PMID:20569134

Constituents of the essential oil of Suregada zanzibariensis leaves are repellent to the mosquito, Anopheles gambiae s.s.

PubMed

Innocent, Ester; Joseph, Cosam C; Gikonyo, Nicholas K; Nkunya, Mayunga H H; Hassanali, Ahmed

2010-01-01

In traditional African communities, repellent volatiles from certain plants generated by direct burning or by thermal expulsion have played an important role in protecting households against vectors of malaria and other diseases. Previous research on volatile constituents of plants has shown that some are good sources of potent mosquito repellents. In this bioprospecting initiative, the essential oil of leaves of the tree, Suregada zanzibariensis Verdc. (Angiospermae: Euphobiaceae) was tested against the mosquito, Anopheles gambiae s.s. Giles (Diptera: Culicidae) and found to be repellent. Gas chromatography (GC), GC-linked mass spectrometry (GC-MS) and, where possible, GC-co-injections with authentic compounds, led to the identification of about 34 compounds in the essential oil. About 56% of the constituents were terpenoid ketones, mostly methyl ketones. Phenylacetaldehyde (14.4%), artemisia ketone (10.1%), (1S)-(-)-verbenone (12.1%) and geranyl acetone (9.4%) were the main constituents. Apart from phenylacetaldehyde, repellent activities of the other main constituents were higher than that of the essential oil. The blends of the main constituents in proportions found in the essential oil were more repellent to An. gambiae s.s. than was the parent oil (p < 0.05), and the presence of artemisia ketone in the blend caused a significant increase in the repellency of the resulting blend. These results suggested that blends of some terpenoid ketones can serve as effective An. gambiae s.s. mosquito repellents.

A highly conserved candidate chemoreceptor expressed in both olfactory and gustatory tissues in the malaria vector Anopheles gambiae.

PubMed

Pitts, R Jason; Fox, A Nicole; Zwiebel, Laurence J

2004-04-06

Anopheles gambiae is a highly anthropophilic mosquito responsible for the majority of malaria transmission in Africa. The biting and host preference behavior of this disease vector is largely influenced by its sense of smell, which is presumably facilitated by G protein-coupled receptor signaling [Takken, W. & Knols, B. (1999) Annu. Rev. Entomol. 44, 131-157]. Because of the importance of host preference to the mosquitoes' ability to transmit disease, we have initiated studies intended to elucidate the molecular mechanisms underlying olfaction in An. gambiae. In the course of these studies, we have identified a number of genes potentially involved in signal transduction, including a family of candidate odorant receptors. One of these receptors, encoded by GPRor7 (hereafter referred to as AgOr7), is remarkably similar to an odorant receptor that is expressed broadly in olfactory tissues and has been identified in Drosophila melanogaster and other insects [Krieger, J., Klink, O., Mohl, C., Raming, K. & Breer, H. (2003) J. Comp. Physiol. A 189, 519-526; Vosshall, L. B., Amrein, H., Morozov, P. S., Rzhetsky, A. & Axel, R. (1999) Cell 96, 725-736]. We have observed AgOr7 expression in olfactory and gustatory tissues in adult An. gambiae and during several stages of the mosquitoes' development. Within the female adult peripheral chemosensory system, antiserum against the AgOR7 polypeptide labels most sensilla of the antenna and maxillary palp as well as a subset of proboscis sensilla. Furthermore, AgOR7 antiserum labeling is observed within the larval antenna and maxillary palpus. These results are consistent with a role for AgOr7 in both olfaction and gustation in An. gambiae and raise the possibility that AgOr7 orthologs may also be of general importance to both modalities of chemosensation in other insects.

Inversion 2La is associated with enhanced desiccation resistance in Anopheles gambiae.

PubMed

Gray, Emilie M; Rocca, Kyle A C; Costantini, Carlo; Besansky, Nora J

2009-09-21

Anopheles gambiae, the principal vector of malignant malaria in Africa, occupies a wide range of habitats. Environmental flexibility may be conferred by a number of chromosomal inversions non-randomly associated with aridity, including 2La. The purpose of this study was to determine the physiological mechanisms associated with the 2La inversion that may result in the preferential survival of its carriers in hygrically-stressful environments. Two homokaryotypic populations of A. gambiae (inverted 2La and standard 2L+(a)) were created from a parental laboratory colony polymorphic for 2La and standard for all other known inversions. Desiccation resistance, water, energy and dry mass of adult females of both populations were compared at several ages and following acclimation to a more arid environment. Females carrying 2La were significantly more resistant to desiccation than 2L+(a) females at emergence and four days post-emergence, for different reasons. Teneral 2La females had lower rates of water loss than their 2L+(a) counterparts, while at four days, 2La females had higher initial water content. No differences in desiccation resistance were found at eight days, with or without acclimation. However, acclimation resulted in both populations significantly reducing their rates of water loss and increasing their desiccation resistance. Acclimation had contrasting effects on the body characteristics of the two populations: 2La females boosted their glycogen stores and decreased lipids, whereas 2La females did the contrary. Variation in rates of water loss and response to acclimation are associated with alternative arrangements of the 2La inversion. Understanding the mechanisms underlying these traits will help explain how inversion polymorphisms permit exploitation of a heterogeneous environment by this disease vector.

Engineered single nucleotide polymorphisms in the mosquito MEK docking site alter Plasmodium berghei development in Anopheles gambiae.

PubMed

Brenton, Ashley A; Souvannaseng, Lattha; Cheung, Kong; Anishchenko, Michael; Brault, Aaron C; Luckhart, Shirley

2014-06-23

Susceptibility to Plasmodium infection in Anopheles gambiae has been proposed to result from naturally occurring polymorphisms that alter the strength of endogenous innate defenses. Despite the fact that some of these mutations are known to introduce non-synonymous substitutions in coding sequences, these mutations have largely been used to rationalize knockdown of associated target proteins to query the effects on parasite development in the mosquito host. Here, we assay the effects of engineered mutations on an immune signaling protein target that is known to control parasite sporogonic development. By this proof-of-principle work, we have established that naturally occurring mutations can be queried for their effects on mosquito protein function and on parasite development and that this important signaling pathway can be genetically manipulated to enhance mosquito resistance. We introduced SNPs into the A. gambiae MAPK kinase MEK to alter key residues in the N-terminal docking site (D-site), thus interfering with its ability to interact with the downstream kinase target ERK. ERK phosphorylation levels in vitro and in vivo were evaluated to confirm the effects of MEK D-site mutations. In addition, overexpression of various MEK D-site alleles was used to assess P. berghei infection in A. gambiae. The MEK D-site contains conserved lysine residues predicted to mediate protein-protein interaction with ERK. As anticipated, each of the D-site mutations (K3M, K6M) suppressed ERK phosphorylation and this inhibition was significant when both mutations were present. Tissue-targeted overexpression of alleles encoding MEK D-site polymorphisms resulted in reduced ERK phosphorylation in the midgut of A. gambiae. Furthermore, as expected, inhibition of MEK-ERK signaling due to D-site mutations resulted in reduction in P. berghei development relative to infection in the presence of overexpressed catalytically active MEK. MEK-ERK signaling in A. gambiae, as in model organisms and

CYP6 P450 enzymes and ACE-1 duplication produce extreme and multiple insecticide resistance in the malaria mosquito Anopheles gambiae.

PubMed

Edi, Constant V; Djogbénou, Luc; Jenkins, Adam M; Regna, Kimberly; Muskavitch, Marc A T; Poupardin, Rodolphe; Jones, Christopher M; Essandoh, John; Kétoh, Guillaume K; Paine, Mark J I; Koudou, Benjamin G; Donnelly, Martin J; Ranson, Hilary; Weetman, David

2014-03-01

Malaria control relies heavily on pyrethroid insecticides, to which susceptibility is declining in Anopheles mosquitoes. To combat pyrethroid resistance, application of alternative insecticides is advocated for indoor residual spraying (IRS), and carbamates are increasingly important. Emergence of a very strong carbamate resistance phenotype in Anopheles gambiae from Tiassalé, Côte d'Ivoire, West Africa, is therefore a potentially major operational challenge, particularly because these malaria vectors now exhibit resistance to multiple insecticide classes. We investigated the genetic basis of resistance to the most commonly-applied carbamate, bendiocarb, in An. gambiae from Tiassalé. Geographically-replicated whole genome microarray experiments identified elevated P450 enzyme expression as associated with bendiocarb resistance, most notably genes from the CYP6 subfamily. P450s were further implicated in resistance phenotypes by induction of significantly elevated mortality to bendiocarb by the synergist piperonyl butoxide (PBO), which also enhanced the action of pyrethroids and an organophosphate. CYP6P3 and especially CYP6M2 produced bendiocarb resistance via transgenic expression in Drosophila in addition to pyrethroid resistance for both genes, and DDT resistance for CYP6M2 expression. CYP6M2 can thus cause resistance to three distinct classes of insecticide although the biochemical mechanism for carbamates is unclear because, in contrast to CYP6P3, recombinant CYP6M2 did not metabolise bendiocarb in vitro. Strongly bendiocarb resistant mosquitoes also displayed elevated expression of the acetylcholinesterase ACE-1 gene, arising at least in part from gene duplication, which confers a survival advantage to carriers of additional copies of resistant ACE-1 G119S alleles. Our results are alarming for vector-based malaria control. Extreme carbamate resistance in Tiassalé An. gambiae results from coupling of over-expressed target site allelic variants with

Living at the edge: biogeographic patterns of habitat segregation conform to speciation by niche expansion in Anopheles gambiae

PubMed Central

Costantini, Carlo; Ayala, Diego; Guelbeogo, Wamdaogo M; Pombi, Marco; Some, Corentin Y; Bassole, Imael HN; Ose, Kenji; Fotsing, Jean-Marie; Sagnon, N'Falé; Fontenille, Didier; Besansky, Nora J; Simard, Frédéric

2009-01-01

Background Ongoing lineage splitting within the African malaria mosquito Anopheles gambiae is compatible with ecological speciation, the evolution of reproductive isolation by divergent natural selection acting on two populations exploiting alternative resources. Divergence between two molecular forms (M and S) identified by fixed differences in rDNA, and characterized by marked, although incomplete, reproductive isolation is occurring in West and Central Africa. To elucidate the role that ecology and geography play in speciation, we carried out a countrywide analysis of An. gambiae M and S habitat requirements, and that of their chromosomal variants, across Burkina Faso. Results Maps of relative abundance by geostatistical interpolators produced a distinct pattern of distribution: the M-form dominated in the northernmost arid zones, the S-form in the more humid southern regions. Maps of habitat suitability, quantified by Ecological Niche Factor Analysis based on 15 eco-geographical variables revealed less contrast among forms. M was peculiar as it occurred proportionally more in habitat of marginal quality. Measures of ecological niche breadth and overlap confirmed the mismatch between the fundamental and realized patterns of habitat occupation: forms segregated more than expected from the extent of divergence of their environmental envelope – a signature of niche expansion. Classification of chromosomal arm 2R karyotypes by multilocus genetic clustering identified two clusters loosely corresponding to molecular forms, with 'mismatches' representing admixed individuals due to shared ancestral polymorphism and/or residual hybridization. In multivariate ordination space, these karyotypes plotted in habitat of more marginal quality compared to non-admixed, 'typical', karyotypes. The distribution of 'typical' karyotypes along the main eco-climatic gradient followed a consistent pattern within and between forms, indicating an adaptive role of inversions at this

Olyset Duo® (a Pyriproxyfen and Permethrin Mixture Net): An Experimental Hut Trial against Pyrethroid Resistant Anopheles gambiae and Culex quinquefasciatus in Southern Benin

PubMed Central

Ngufor, Corine; N’Guessan, Raphael; Fagbohoun, Josias; Odjo, Abibatou; Malone, David; Akogbeto, Martin; Rowland, Mark

2014-01-01

Background Alternative compounds which can complement pyrethroids on long-lasting insecticidal nets (LN) in the control of pyrethroid resistant malaria vectors are urgently needed. Pyriproxyfen (PPF), an insect growth regulator, reduces the fecundity and fertility of adult female mosquitoes. LNs containing a mixture of pyriproxyfen and pyrethroid could provide personal protection through the pyrethroid component and reduce vector abundance in the next generation through the sterilizing effect of pyriproxyfen. Method The efficacy of Olyset Duo, a newly developed mixture LN containing pyriproxyfen and permethrin, was evaluated in experimental huts in southern Benin against pyrethroid resistant Anopheles gambiae and Culex quinquefasciatus. Comparison was made with Olyset Net® (permethrin alone) and a LN with pyriproxyfen alone (PPF LN). Laboratory tunnel tests were performed to substantiate the findings in the experimental huts. Results Overall mortality of wild pyrethroid resistant An. gambiae s.s. was significantly higher with Olyset Duo than with Olyset Net (50% vs. 27%, P = 0.01). Olyset DUO was more protective than Olyset Net (71% vs. 3%, P<0.001). The oviposition rate of surviving blood-fed An. gambiae from the control hut was 37% whereas none of those from Olyset Duo and PPF LN huts laid eggs. The tunnel test results were consistent with the experimental hut results. Olyset Duo was more protective than Olyset Net in the huts against wild pyrethroid resistant Cx. quinquefasciatus although mortality rates of this species did not differ significantly between Olyset Net and Olyset Duo. There was no sterilizing effect on surviving blood-fed Cx. quinquefasciatus with the PPF-treated nets. Conclusion Olyset Duo was superior to Olyset Net in terms of personal protection and killing of pyrethroid resistant An. gambiae, and sterilized surviving blood-fed mosquitoes. Mixing pyrethroid and pyriproxyfen on a LN shows potential for malaria control and management of

A High Affinity Adenosine Kinase from Anopheles gambiae

PubMed Central

Cassera, María B.; Ho, Meng-Chiao; Merino, Emilio F.; Burgos, Emmanuel S.; Rinaldo-Matthis, Agnes; Almo, Steven C.; Schramm, Vern L.

2011-01-01

Genome analysis revealed a mosquito orthologue of adenosine kinase in Anopheles gambiae (AgAK; the most important vector for the transmission of Plasmodium falciparum in Africa). P. falciparum are purine auxotrophs and do not express an adenosine kinase but rely on their hosts for purines. AgAK was kinetically characterized and found to have the highest affinity for adenosine (Km 8.1 nM) of any known adenosine kinase. AgAK is specific for adenosine at the nucleoside site but several nucleotide triphosphate phosphoryl donors are tolerated. The AgAK crystal structure with a bound bisubstrate analogue Ap4A (2.0 Å resolution) reveals interactions for adenosine, ATP and the geometry for phosphoryl transfer. The polyphosphate charge is partly neutralized by a bound Mg2+ ion and an ion pair to a catalytic site Arg. The AgAK structure consists of a large catalytic core in a three-layered α/β/α sandwich, and a small cap domain in contact with adenosine. The specificity and tight-binding for adenosine arises from hydrogen bond interactions of Asn14, Leu16, Leu40, Leu133, Leu168, Phe168 and Thr171 and the backbone of Ile39 and Phe168 with the adenine ring as well as through hydrogen bond interactions between Asp18, Gly64 and Asn68 and the ribosyl 2′- and 3′-hydroxyl groups. The structure is more similar to human adenosine kinase (48% identity) than to AK from Toxoplasma gondii (31% identity). With this extraordinary affinity for AgAK, adenosine is efficiently captured and converted to AMP at near the diffusion limit, suggesting an important role of this enzyme to maintain the adenine nucleotide pool. mRNA analysis verifies that AgAK transcripts are produced in the adult insects. PMID:21247194

A High-Affinity Adenosine Kinase from Anopheles Gambiae

DOE Office of Scientific and Technical Information (OSTI.GOV)

M Cassera; M Ho; E Merino

2011-12-31

Genome analysis revealed a mosquito orthologue of adenosine kinase in Anopheles gambiae (AgAK; the most important vector for the transmission of Plasmodium falciparum in Africa). P. falciparum are purine auxotrophs and do not express an adenosine kinase but rely on their hosts for purines. AgAK was kinetically characterized and found to have the highest affinity for adenosine (K{sub m} = 8.1 nM) of any known adenosine kinase. AgAK is specific for adenosine at the nucleoside site, but several nucleotide triphosphate phosphoryl donors are tolerated. The AgAK crystal structure with a bound bisubstrate analogue Ap{sub 4}A (2.0 {angstrom} resolution) reveals interactionsmore » for adenosine and ATP and the geometry for phosphoryl transfer. The polyphosphate charge is partly neutralized by a bound Mg{sup 2+} ion and an ion pair to a catalytic site Arg. The AgAK structure consists of a large catalytic core in a three-layer {alpha}/{beta}/{alpha} sandwich, and a small cap domain in contact with adenosine. The specificity and tight binding for adenosine arise from hydrogen bond interactions of Asn14, Leu16, Leu40, Leu133, Leu168, Phe168, and Thr171 and the backbone of Ile39 and Phe168 with the adenine ring as well as through hydrogen bond interactions between Asp18, Gly64, and Asn68 and the ribosyl 2'- and 3'-hydroxyl groups. The structure is more similar to that of human adenosine kinase (48% identical) than to that of AK from Toxoplasma gondii (31% identical). With this extraordinary affinity for AgAK, adenosine is efficiently captured and converted to AMP at near the diffusion limit, suggesting an important role for this enzyme in the maintenance of the adenine nucleotide pool. mRNA analysis verifies that AgAK transcripts are produced in the adult insects.« less

Effects of available sugar on the reproductive fitness and vectorial capacity of the malaria vector Anopheles gambiae (Diptera: Culicidae).

PubMed

Gary, R E; Foster, W A

2001-01-01

Although females of most mosquito species are known to use sugar as a necessary source of energy, female Anopheles gambiae Giles sensu stricto are thought to use it facultatively or not at all. However, field evidence of sugar-free living is inconclusive, and the implications for reproductive fitness and vectorial capacity are unknown. To evaluate the role that sugar may play in the ecology of these mosquitoes, mated female An. gambiae in the laboratory were given access to either no food (water only), 10% sucrose, human blood, or human blood + 10% sucrose, and comparisons of daily mortality, fecundity, and biting frequency were made. The effect of sugar availability on vectorial capacity and the intrinsic rate of increase, a measure of fitness, then were determined. Females (pooled and individual) given blood + sugar lived significantly longer than did those on the other diets. Daily fecundity was higher for females given blood alone than for those fed blood + sugar (13 versus 9 eggs per female daily). However, total fecundity and intrinsic rate of increase were not affected by sugar availability. Biting frequency was significantly higher (0.41 versus 0.26 bites per female per day) for females given blood alone. Despite the reduced survivorship, exclusive blood-feeding led to a theoretically higher vectorial capacity for Plasmodium falciparum at 27 degrees C. These data indicate that female An. gambiae could replace sugar with increased blood feeding without suppressing reproductive fitness. Increased blood feeding could, in turn, increase the rate of malaria transmission and may explain the unusual efficiency of this vector.

Regulation of carbohydrate metabolism and flight performance by a hypertrehalosaemic hormone in the mosquito Anopheles gambiae

PubMed Central

Kaufmann, Christian; Brown, Mark R.

2008-01-01

The role of adipokinetic hormones (AKHs) in the regulation of carbohydrate and lipid metabolism and flight performance was evaluated for females of the African malaria mosquito, Anopheles gambiae. Injection of various dosages of synthetic Anoga-AKH-I increased carbohydrate levels in the haemolymph and reduced glycogen reserves in sugar-fed females but did not affect lipid levels. Anoga-AKH-I enhanced the flight performance of both intact and decapitated sugar-fed females, during a 4 hour flight period. Anoga-AKH-II had no effect on carbohydrate or lipid levels or flight performance, thus its function remains unknown. Targeted RNA-interference lowered Anoga-AKH receptor expression in sugar-fed females, consequently injections of Anoga-AKH-I failed to mobilize glycogen reserves. Taken together, these results show that a primary role for the neurohormone, Anoga-AKH-I, is to elevate trehalose levels in the haemolymph of female mosquitoes. PMID:18062987

A behavioral mechanism underlying ecological divergence in the malaria mosquito Anopheles gambiae.

PubMed

Gimonneau, Geoffrey; Bouyer, Jérémy; Morand, Serge; Besansky, Nora J; Diabate, Abdoulaye; Simard, Frédéric

2010-09-01

Disruptive selection mediated by predation on aquatic immature stages has been proposed as a major force driving ecological divergence and fostering speciation between the M and S molecular forms of the African malaria mosquito, Anopheles gambiae. In the dry savannahs of West Africa where both molecular forms co-occur, the S form thrives in temporary pools filled with rainwater, whereas the M form preferentially breeds in permanent freshwater habitats where predator pressure is higher. Here, we explored the proximal mechanisms by which predation may contribute to habitat segregation between molecular forms using progeny of female mosquitoes captured in Burkina Faso. We show that the S form suffers higher predation rates than the M form when simultaneously exposed to the widespread predator, Anisops jaczewskii in an experimental arena. Furthermore, behavioral plasticity induced by exposure to the predator was observed in the M form, but not in the S form, and may partially explain its habitat use and ecological divergence from the S form. We discuss the role of adaptive phenotypic plasticity in allowing successful colonization of a new ecological niche by the M form and highlight further research areas that need to be addressed for a better understanding of the ultimate mechanisms underlying ecological speciation in this pest of major medical importance.

Microbial larvicides for malaria control in The Gambia

PubMed Central

Majambere, Silas; Lindsay, Steven W; Green, Clare; Kandeh, Balla; Fillinger, Ulrike

2007-01-01

Background Mosquito larval control may prove to be an effective tool for incorporating into integrated vector management (IVM) strategies for reducing malaria transmission. Here the efficacy of microbial larvicides against Anopheles gambiae s.l. was tested in preparation for a large-scale larviciding programme in The Gambia. Methods The impact of water-dispersible (WDG) and corn granule (CG) formulations of commercial Bacillus sphaericus strain 2362 (Bs; VectoLex®) and Bacillus thuringiensis var.israelensis strain AM65-52 (Bti; VectoBac®) on larval development were tested under laboratory and field conditions to (1) identify the susceptibility of local vectors, (2) evaluate the residual effect and re-treatment intervals, (3) test the effectiveness of the microbials under operational application conditions and (4) develop a method for large-scale application. Results The major malaria vectors were highly susceptible to both microbials. The lethal concentration (LC) to kill 95% of third instar larvae of Anopheles gambiae s.s. after 24 hours was 0.023 mg/l (14.9 BsITU/l) for Bs WDG and 0.132 mg/l (396 ITU/l) for Bti WDG. In general Bs had little residual effect under field conditions even when the application rate was 200 times greater than the LC95. However, there was a residual effect up to 10 days in standardized field tests implemented during the dry season. Both microbials achieved 100% mortality of larvae 24–48 hours post-application but late instar larvae were detected 4 days after treatment. Pupae development was reduced by 94% (95% Confidence Interval = 90.8–97.5%) at weekly re-treatment intervals. Field tests showed that Bs had no residual activity against anopheline larvae. Both microbials provided complete protection when applied weekly. The basic training of personnel in identification of habitats, calibration of application equipment and active larviciding proved to be successful and achieved full coverage and control of mosquito larvae for three

The physicochemical and environmental factors affecting the distribution of Anopheles merus along the Kenyan coast.

PubMed

Kipyab, Pamela C; Khaemba, Battan M; Mwangangi, Joseph M; Mbogo, Charles M

2015-04-11

Members of the Anopheles gambiae complex are the main transmitters of malaria. Anopheles merus is a member of the complex found along the Kenyan coast because it breeds in saline waters. An entomological study was conducted in Garithe Malindi District, to investigate the physicochemical and environmental factors affecting the distribution of An. merus. Field and laboratory studies were used to investigate the breeding habitats of the subspecies. Mosquito larvae were sampled using standard dipping technique from small pockets of pools, ponds, hoof prints, road drain, wells and mangrove swamps found in Garithe. All 3(rd) and 4(th) instars of Anopheles larvae sampled were identified microscopically into species. A representative of Anopheles gambiae complex was then identified to specific sibling species using r-DNA PCR technique. The habitats were characterized based on temperature, conductivity, salinity, dissolved oxygen, total dissolved solids, pH, size, distance to nearest house, canopy coverage, surface debris, presence of algae, emergent plants, turbidity and habitat types. A total of 159 morphologically identified late stage instar Anopheles gambiae s.l larvae were selected for r-DNA analysis by PCR. Out of these, 60.4% (n = 96) were Anopheles merus, 8.8% (n = 14) were Anopheles arabiensis, 18.2% (n = 29) were Anopheles gambiae s.s and 12.6% (n = 20) were unknown. Using paired t-test (t (121) = -3.331, P = 0.001) a significantly high proportion of An. merus was observed in all habitats compared to An. arabiensis, and An. gambiae s. s. In habitat characterization, Pearson's correlation analysis test showed different parameters being associated with the occurrence of An. merus larvae in the different habitats sampled. Six out of the 55 correlation coefficients (10.9%) were statistically significant, suggesting non-random association between some pairs of variables. Those that had a significantly high positive correlation with An. merus

PCR-based karyotyping of Anopheles gambiae inversion 2Rj identifies the BAMAKO chromosomal form.

PubMed

Coulibaly, Mamadou B; Pombi, Marco; Caputo, Beniamino; Nwakanma, Davis; Jawara, Musa; Konate, Lassana; Dia, Ibrahima; Fofana, Abdrahamane; Kern, Marcia; Simard, Frédéric; Conway, David J; Petrarca, Vincenzo; della Torre, Alessandra; Traoré, Sékou; Besansky, Nora J

2007-10-01

The malaria vector Anopheles gambiae is polymorphic for chromosomal inversions on the right arm of chromosome 2 that segregate nonrandomly between assortatively mating populations in West Africa. One such inversion, 2Rj, is associated with the BAMAKO chromosomal form endemic to southern Mali and northern Guinea Conakry near the Niger River. Although it exploits a unique ecology and both molecular and chromosomal data suggest reduced gene flow between BAMAKO and other A. gambiae populations, no molecular markers exist to identify this form. To facilitate study of the BAMAKO form, a PCR assay for molecular karyotyping of 2Rj was developed based on sequences at the breakpoint junctions. The assay was extensively validated using more than 700 field specimens whose karyotypes were determined in parallel by cytogenetic and molecular methods. As inversion 2Rj also occurs in SAVANNA populations outside the geographic range of BAMAKO, samples were tested from Senegal, Cameroon and western Guinea Conakry as well as from Mali. In southern Mali, where 2Rj polymorphism in SAVANNA populations was very low and most of the 2Rj homozygotes were found in BAMAKO karyotypes, the molecular and cytogenetic methods were almost perfectly congruent. Elsewhere agreement between the methods was much poorer, as the molecular assay frequently misclassified 2Rj heterozygotes as 2R+j standard homozygotes. Molecular karyotyping of 2Rj is robust and accurate on 2R+j standard and 2Rj inverted homozygotes. Therefore, the proposed approach overcomes the lack of a rapid tool for identifying the BAMAKO form across developmental stages and sexes, and opens new perspectives for the study of BAMAKO ecology and behaviour. On the other hand, the method should not be applied for molecular karyotyping of j-carriers within the SAVANNA chromosomal form.

Analysis of near infrared spectra for age-grading of wild populations of Anopheles gambiae.

PubMed

Krajacich, Benjamin J; Meyers, Jacob I; Alout, Haoues; Dabiré, Roch K; Dowell, Floyd E; Foy, Brian D

2017-11-07

Understanding the age-structure of mosquito populations, especially malaria vectors such as Anopheles gambiae, is important for assessing the risk of infectious mosquitoes, and how vector control interventions may impact this risk. The use of near-infrared spectroscopy (NIRS) for age-grading has been demonstrated previously on laboratory and semi-field mosquitoes, but to date has not been utilized on wild-caught mosquitoes whose age is externally validated via parity status or parasite infection stage. In this study, we developed regression and classification models using NIRS on datasets of wild An. gambiae (s.l.) reared from larvae collected from the field in Burkina Faso, and two laboratory strains. We compared the accuracy of these models for predicting the ages of wild-caught mosquitoes that had been scored for their parity status as well as for positivity for Plasmodium sporozoites. Regression models utilizing variable selection increased predictive accuracy over the more common full-spectrum partial least squares (PLS) approach for cross-validation of the datasets, validation, and independent test sets. Models produced from datasets that included the greatest range of mosquito samples (i.e. different sampling locations and times) had the highest predictive accuracy on independent testing sets, though overall accuracy on these samples was low. For classification, we found that intramodel accuracy ranged between 73.5-97.0% for grouping of mosquitoes into "early" and "late" age classes, with the highest prediction accuracy found in laboratory colonized mosquitoes. However, this accuracy was decreased on test sets, with the highest classification of an independent set of wild-caught larvae reared to set ages being 69.6%. Variation in NIRS data, likely from dietary, genetic, and other factors limits the accuracy of this technique with wild-caught mosquitoes. Alternative algorithms may help improve prediction accuracy, but care should be taken to either maximize

The Anopheles gambiae 2La chromosome inversion is associated with susceptibility to Plasmodium falciparum in Africa

PubMed Central

Riehle, Michelle M; Bukhari, Tullu; Gneme, Awa; Guelbeogo, Wamdaogo M; Coulibaly, Boubacar; Fofana, Abdrahamane; Pain, Adrien; Bischoff, Emmanuel; Renaud, Francois; Beavogui, Abdoul H; Traore, Sekou F; Sagnon, N’Fale; Vernick, Kenneth D

2017-01-01

Chromosome inversions suppress genetic recombination and establish co-adapted gene complexes, or supergenes. The 2La inversion is a widespread polymorphism in the Anopheles gambiae species complex, the major African mosquito vectors of human malaria. Here we show that alleles of the 2La inversion are associated with natural malaria infection levels in wild-captured vectors from West and East Africa. Mosquitoes carrying the more-susceptible allele (2L+a) are also behaviorally less likely to be found inside houses. Vector control tools that target indoor-resting mosquitoes, such as bednets and insecticides, are currently the cornerstone of malaria control in Africa. Populations with high levels of the 2L+a allele may form reservoirs of persistent outdoor malaria transmission requiring novel measures for surveillance and control. The 2La inversion is a major and previously unappreciated component of the natural malaria transmission system in Africa, influencing both malaria susceptibility and vector behavior. DOI: http://dx.doi.org/10.7554/eLife.25813.001 PMID:28643631

The Anopheles gambiae 2La chromosome inversion is associated with susceptibility to Plasmodium falciparum in Africa.

PubMed

Riehle, Michelle M; Bukhari, Tullu; Gneme, Awa; Guelbeogo, Wamdaogo M; Coulibaly, Boubacar; Fofana, Abdrahamane; Pain, Adrien; Bischoff, Emmanuel; Renaud, Francois; Beavogui, Abdoul H; Traore, Sekou F; Sagnon, N'Fale; Vernick, Kenneth D

2017-06-23

Chromosome inversions suppress genetic recombination and establish co-adapted gene complexes, or supergenes. The 2La inversion is a widespread polymorphism in the Anopheles gambiae species complex, the major African mosquito vectors of human malaria. Here we show that alleles of the 2La inversion are associated with natural malaria infection levels in wild-captured vectors from West and East Africa. Mosquitoes carrying the more-susceptible allele (2L+ a ) are also behaviorally less likely to be found inside houses. Vector control tools that target indoor-resting mosquitoes, such as bednets and insecticides, are currently the cornerstone of malaria control in Africa. Populations with high levels of the 2L+ a allele may form reservoirs of persistent outdoor malaria transmission requiring novel measures for surveillance and control. The 2La inversion is a major and previously unappreciated component of the natural malaria transmission system in Africa, influencing both malaria susceptibility and vector behavior.

Expression of the cytochrome P450s, CYP6P3 and CYP6M2 are significantly elevated in multiple pyrethroid resistant populations of Anopheles gambiae s.s. from Southern Benin and Nigeria

PubMed Central

Djouaka, Rousseau F; Bakare, Adekunle A; Coulibaly, Ousmane N; Akogbeto, Martin C; Ranson, Hilary; Hemingway, Janet; Strode, Clare

2008-01-01

Background Insecticide resistance in Anopheles mosquitoes is threatening the success of malaria control programmes. This is particularly true in Benin where pyrethroid resistance has been linked to the failure of insecticide treated bed nets. The role of mutations in the insecticide target sites in conferring resistance has been clearly established. In this study, the contribution of other potential resistance mechanisms was investigated in Anopheles gambiae s.s. from a number of localities in Southern Benin and Nigeria. The mosquitoes were sampled from a variety of breeding sites in a preliminary attempt to investigate the role of contamination of mosquito breeding sites in selecting for resistance in adult mosquitoes. Results All mosquitoes sampled belonged to the M form of An. gambiae s.s. There were high levels of permethrin resistance in an agricultural area (Akron) and an urban area (Gbedjromede), low levels of resistance in mosquito samples from an oil contaminated site (Ojoo) and complete susceptibility in the rural Orogun location. The target site mutation kdrW was detected at high levels in two of the populations (Akron f = 0.86 and Gbedjromede f = 0.84) but was not detected in Ojoo or Orogun. Microarray analysis using the Anopheles gambiae detox chip identified two P450s, CYP6P3 and CYP6M2 up regulated in all three populations, the former was expressed at particularly high levels in the Akron (12.4-fold) and Ojoo (7.4-fold) populations compared to the susceptible population. Additional detoxification and redox genes were also over expressed in one or more populations including two cuticular pre-cursor genes which were elevated in two of the three resistant populations. Conclusion Multiple resistance mechanisms incurred in the different breeding sites contribute to resistance to permethrin in Benin. The cytochrome P450 genes, CYP6P3 and CYP6M2 are upregulated in all three resistant populations analysed. Several additional potential resistance mechanisms

Comparison of the Insecticidal Characteristics of Commercially Available Plant Essential Oils Against Aedes aegypti and Anopheles gambiae (Diptera: Culicidae).

PubMed

Norris, Edmund J; Gross, Aaron D; Dunphy, Brendan M; Bessette, Steven; Bartholomay, Lyric; Coats, Joel R

2015-09-01

Aedes aegypti and Anopheles gambiae are two mosquito species that represent significant threats to global public health as vectors of Dengue virus and malaria parasites, respectively. Although mosquito populations have been effectively controlled through the use of synthetic insecticides, the emergence of widespread insecticide-resistance in wild mosquito populations is a strong motivation to explore new insecticidal chemistries. For these studies, Ae. aegypti and An. gambiae were treated with commercially available plant essential oils via topical application. The relative toxicity of each essential oil was determined, as measured by the 24-h LD(50) and percentage knockdown at 1 h, as compared with a variety of synthetic pyrethroids. For Ae. aegypti, the most toxic essential oil (patchouli oil) was ∼1,700-times less toxic than the least toxic synthetic pyrethroid, bifenthrin. For An. gambiae, the most toxic essential oil (patchouli oil) was ∼685-times less toxic than the least toxic synthetic pyrethroid. A wide variety of toxicities were observed among the essential oils screened. Also, plant essential oils were analyzed via gas chromatography/mass spectrometry (GC/MS) to identify the major components in each of the samples screened in this study. While the toxicities of these plant essential oils were demonstrated to be lower than those of the synthetic pyrethroids tested, the large amount of GC/MS data and bioactivity data for each essential oil presented in this study will serve as a valuable resource for future studies exploring the insecticidal quality of plant essential oils. © The Authors 2015. Published by Oxford University Press on behalf of Entomological Society of America. All rights reserved. For Permissions, please email: journals.permissions@oup.com.

Larvicidal effect of disinfectant soap on Anopheles gambiae s.s (Diptera: Culicidae) in laboratory and semifield environs.

PubMed

Mdoe, France P; Nkwengulila, Gamba; Chobu, Mariam; Lyaruu, Lucile; Gyunda, Israel L; Mbepera, Saada; Xue, Rui-De; Kweka, Eliningaya J

2014-05-03

Mosquito larval control using chemicals and biological agents is of paramount importance in vector population and disease incidence reduction. A commercial synthetic disinfectant soap was evaluated against larvae of Anopheles gambiae s.s. in both laboratory and semi field conditions. Five concentrations of commercial synthetic disinfectant soap (0.0001, 0.001, 0.01, 0.1 and 1%) were prepared and evaluated against third instar larvae in laboratory and semi field environments. Mortality was scored at 12, 24, 48, and 72 hrs. Each dosage had 6 replicates, having twenty 3rd instar larvae of An.gambiae s.s. In the laboratory phase, all dosages had significantly higher larval mortalities than in controls, while in semi field conditions, the dosages of 0.0001, 0.001 and 0.01% had lower mortalities than laboratory trials. In the comparison between semi field and laboratory trials, only 0.1 and 1% dosage had significant difference with more mortality in semifield conditions. Proportions of larvae that died during mortality monitoring intervals in laboratory and semi field had significant differences only at 12 hrs and 72 hrs. The findings of this study have demonstrated that the mortality of larvae caused by commercial synthetic disinfectant soap is worth further studies in open water bodies. More studies are necessary to find out the effect of sunlight on the chemistry of the synthetic disinfectant and other variables in small scale full field trials.

A spatial individual-based model predicting a great impact of copious sugar sources and resting sites on survival of Anopheles gambiae and malaria parasite transmission.

PubMed

Zhu, Lin; Qualls, Whitney A; Marshall, John M; Arheart, Kris L; DeAngelis, Donald L; McManus, John W; Traore, Sekou F; Doumbia, Seydou; Schlein, Yosef; Müller, Günter C; Beier, John C

2015-02-05

Agent-based modelling (ABM) has been used to simulate mosquito life cycles and to evaluate vector control applications. However, most models lack sugar-feeding and resting behaviours or are based on mathematical equations lacking individual level randomness and spatial components of mosquito life. Here, a spatial individual-based model (IBM) incorporating sugar-feeding and resting behaviours of the malaria vector Anopheles gambiae was developed to estimate the impact of environmental sugar sources and resting sites on survival and biting behaviour. A spatial IBM containing An. gambiae mosquitoes and humans, as well as the village environment of houses, sugar sources, resting sites and larval habitat sites was developed. Anopheles gambiae behaviour rules were attributed at each step of the IBM: resting, host seeking, sugar feeding and breeding. Each step represented one second of time, and each simulation was set to run for 60 days and repeated 50 times. Scenarios of different densities and spatial distributions of sugar sources and outdoor resting sites were simulated and compared. When the number of natural sugar sources was increased from 0 to 100 while the number of resting sites was held constant, mean daily survival rate increased from 2.5% to 85.1% for males and from 2.5% to 94.5% for females, mean human biting rate increased from 0 to 0.94 bites per human per day, and mean daily abundance increased from 1 to 477 for males and from 1 to 1,428 for females. When the number of outdoor resting sites was increased from 0 to 50 while the number of sugar sources was held constant, mean daily survival rate increased from 77.3% to 84.3% for males and from 86.7% to 93.9% for females, mean human biting rate increased from 0 to 0.52 bites per human per day, and mean daily abundance increased from 62 to 349 for males and from 257 to 1120 for females. All increases were significant (P < 0.01). Survival was greater when sugar sources were randomly distributed in the whole

Carbamate and Pyrethroid Resistance in the Akron Strain of Anopheles gambiae

PubMed Central

Mutunga, James M.; Anderson, Troy D.; Craft, Derek T.; Gross, Aaron D.; Swale, Daniel R.; Tong, Fan; Wong, Dawn M.; Carlier, Paul R.; Bloomquist, Jeffrey R.

2015-01-01

Insecticide resistance in the malaria vector, Anopheles gambiae is a serious problem, epitomized by the multi-resistant Akron strain, originally isolated in the country of Benin. Here we report resistance in this strain to pyrethroids and DDT (13-fold to 35-fold compared to the susceptible G3 strain), but surprisingly little resistance to etofenprox, a compound sometimes described as a “pseudo-pyrethroid.” There was also strong resistance to topically-applied commercial carbamates (45-fold to 81-fold), except for the oximes aldicarb and methomyl. Biochemical assays showed enhanced cytochrome P450 monooxygenase and carboxylesterase activity, but not that of glutathione-S-transferase. A series of substituted α,α,α,-trifluoroacetophenone oxime methylcarbamates were evaluated for enzyme inhibition potency and toxicity against G3 and Akron mosquitoes. The compound bearing an unsubstituted phenyl ring showed the greatest toxicity to mosquitoes of both strains. Low cross resistance in Akron was retained by all analogs in the series. Kinetic analysis of acetylcholinesterase activity and its inhibition by insecticides in the G3 strain showed inactivation rate constants greater than that of propoxur, and against Akron enzyme inactivation rate constants similar to that of aldicarb. However, inactivation rate constants against recombinant human AChE were essentially identical to that of the G3 strain. Thus, the acetophenone oxime carbamates described here, though potent insecticides that control resistant Akron mosquitoes, require further structural modification to attain acceptable selectivity and human safety. PMID:26047119

The serine protease homolog CLIPA14 modulates the intensity of the immune response in the mosquito Anopheles gambiae

PubMed Central

Nakhleh, Johnny; Christophides, George K.; Osta, Mike A.

2017-01-01

Clip domain serine protease homologs (SPHs) are positive and negative regulators of Anopheles gambiae immune responses mediated by the complement-like protein TEP1 against Plasmodium malaria parasites and other microbial infections. We have previously reported that the SPH CLIPA2 is a negative regulator of the TEP1-mediated response by showing that CLIPA2 knockdown (kd) enhances mosquito resistance to infections with fungi, bacteria, and Plasmodium parasites. Here, we identify another SPH, CLIPA14, as a novel regulator of mosquito immunity. We found that CLIPA14 is a hemolymph protein that is rapidly cleaved following a systemic infection. CLIPA14 kd mosquitoes elicited a potent melanization response against Plasmodium berghei ookinetes and exhibited significantly increased resistance to Plasmodium infections as well as to systemic and oral bacterial infections. The activity of the enzyme phenoloxidase, which initiates melanin biosynthesis, dramatically increased in the hemolymph of CLIPA14 kd mosquitoes in response to systemic bacterial infections. Ookinete melanization and hemolymph phenoloxidase activity were further increased after cosilencing CLIPA14 and CLIPA2, suggesting that these two SPHs act in concert to control the melanization response. Interestingly, CLIPA14 RNAi phenotypes and its infection-induced cleavage were abolished in a TEP1 loss-of-function background. Our results suggest that a complex network of SPHs functions downstream of TEP1 to regulate the melanization reaction. PMID:28928218

Mosquito Protein Kinase G Phosphorylates Flavivirus NS5 and Alters Flight Behavior in Aedes aegypti and Anopheles gambiae

PubMed Central

Keating, Julie A.; Bhattacharya, Dipankar; Rund, Samuel S.C.; Hoover, Spencer; Dasgupta, Ranjit; Lee, Samuel J.; Duffield, Giles E.

2013-01-01

Abstract Many arboviral proteins are phosphorylated in infected mammalian cells, but it is unknown if the same phosphorylation events occur when insects are similarly infected. One of the mammalian kinases responsible for phosphorylation, protein kinase G (PKG), has been implicated in the behavior of multiple nonvector insects, but is unstudied in mosquitoes. PKG from Aedes aegypti was cloned, and phosphorylation of specific viral sites was monitored by mass spectrometry from biochemical and cell culture experiments. PKG from Aedes mosquitoes is able to phosphorylate dengue nonstructural protein 5 (NS5) at specific sites in cell culture and cell-free systems and autophosphorylates its own regulatory domain in a cell-free system. Injecting Aedes aegypti and Anopheles gambiae mosquitoes with a pharmacological PKG activator resulted in increased Aedes wing activity during periods of their natural diurnal/crepuscular activity and increased Anopheles nocturnal locomotor/flight activity. Thus, perturbation of the PKG signaling pathway in mosquitoes alters flight behavior. The demonstrated effect of PKG alterations is consistent with a viral PKG substrate triggering increased PKG activity. This increased PKG activity could be the mechanism by which dengue virus increases flight behavior and possibly facilitates transmission. Whether or not PKG is part of the mechanism by which dengue increases flight behavior, this report is the first to show PKG can modulate behavior in hematophagous disease vectors. PMID:23930976

Interactive cost of Plasmodium infection and insecticide resistance in the malaria vector Anopheles gambiae.

PubMed

Alout, Haoues; Dabiré, Roch K; Djogbénou, Luc S; Abate, Luc; Corbel, Vincent; Chandre, Fabrice; Cohuet, Anna

2016-07-19

Insecticide resistance raises concerns for the control of vector-borne diseases. However, its impact on parasite transmission could be diverse when considering the ecological interactions between vector and parasite. Thus we investigated the fitness cost associated with insecticide resistance and Plasmodium falciparum infection as well as their interactive cost on Anopheles gambiae survival and fecundity. In absence of infection, we observed a cost on fecundity associated with insecticide resistance. However, survival was higher for mosquito bearing the kdr mutation and equal for those with the ace-1(R) mutation compared to their insecticide susceptible counterparts. Interestingly, Plasmodium infection reduced survival only in the insecticide resistant strains but not in the susceptible one and infection was associated with an increase in fecundity independently of the strain considered. This study provides evidence for a survival cost associated with infection by Plasmodium parasite only in mosquito selected for insecticide resistance. This suggests that the selection of insecticide resistance mutation may have disturbed the interaction between parasites and vectors, resulting in increased cost of infection. Considering the fitness cost as well as other ecological aspects of this natural mosquito-parasite combination is important to predict the epidemiological impact of insecticide resistance.

Genetic and environmental factors associated with laboratory rearing affect survival and assortative mating but not overall mating success in Anopheles gambiae sensu stricto.

PubMed

Paton, Doug; Touré, Mahamoudou; Sacko, Adama; Coulibaly, Mamadou B; Traoré, Sékou F; Tripet, Frédéric

2013-01-01

Anopheles gambiae sensu stricto, the main vector of malaria in Africa, is characterized by its vast geographical range and complex population structure. Assortative mating amongst the reproductively isolated cryptic forms that co-occur in many areas poses unique challenges for programs aiming to decrease malaria incidence via the release of sterile or genetically-modified mosquitoes. Importantly, whether laboratory-rearing affects the ability of An. gambiae individuals of a given cryptic taxa to successfully mate with individuals of their own form in field conditions is still unknown and yet crucial for mosquito-releases. Here, the independent effects of genetic and environmental factors associated with laboratory rearing on male and female survival, mating success and assortative mating were evaluated in the Mopti form of An. gambiae over 2010 and 2011. In semi-field enclosures experiments and despite strong variation between years, the overall survival and mating success of male and female progeny from a laboratory strain was not found to be significantly lower than those of the progeny of field females from the same population. Adult progeny from field-caught females reared at the larval stage in the laboratory and from laboratory females reared outdoors exhibited a significant decrease in survival but not in mating success. Importantly, laboratory individuals reared as larvae indoors were unable to mate assortatively as adults, whilst field progeny reared either outdoors or in the laboratory, as well as laboratory progeny reared outdoors all mated significantly assortatively. These results highlight the importance of genetic and environment interactions for the development of An. gambiae's full mating behavioral repertoire and the challenges this creates for mosquito rearing and release-based control strategies.

Experimental hut evaluation of linalool spatial repellent agar gel against Anopheles gambiae sensu stricto mosquitoes in a semi-field system in Bagamoyo, Tanzania.

PubMed

Tambwe, Mgeni Mohamed; Mbeyela, Edgar Mtaki; Massinda, Brian Migamyo; Moore, Sarah Jane; Maia, Marta Ferreira

2014-12-05

Malaria vector control is in need of new tools to face its current challenges such as the spread of pyrethroid-resistance and the increase of outdoor feeding mosquitoes. New strategies such as spatial repellents need to be evaluated as supplemental tools to existing control measures such as insecticide treated bed nets and indoor residual spraying. Linalool is a naturally occurring terpene alcohol commonly found in flowers and spices with reportedly repellent properties. Four experimental huts fitted with exit traps and enclosed inside a large screened semi-field system were used for the evaluation. The tested spatial repellent product consisted of an agar gel emanator containing 73% linalool. Two rounds of experiments using a Latin square design were conducted to evaluate the efficacy of the linalool emanators compared to no treatment (negative control) and a transfluthrin coil (positive) against lab-reared disease free Anopheles gambiae s.s.. The emanators were hung inside experimental huts where two volunteers were sleeping unprotected. The outcome measures were repellency, % feeding inhibition, %mortality and post 24 h % mortality. Unlike the mosquito coil, the linalool emanators did not show any feeding inhibition, repellency or induced mortality compared to the negative control. On the other hand mosquitoes kept for 24 h post exposure were 3 times more likely to die after being exposed to two 73% linalool emanators than the negative control. Our results indicate that linalool agar gel emanators are not adequate as a spatial repellent against Anopheles gambiae s.s.. However adding linalool to known repellent formulations could be advantageous, not only because of its pleasant scent but also because of the delayed mortality effect it has on mosquitoes.

Crepuscular Behavioral Variation and Profiling of Opsin Genes in Anopheles gambiae and Anopheles stephensi (Diptera: Culicidae)

PubMed Central

Jenkins, Adam M.; Muskavitch, Marc A. T.

2015-01-01

We understand little about photopreference and the molecular mechanisms governing vision-dependent behavior in vector mosquitoes. Investigations of the influence of photopreference on adult mosquito behaviors such as endophagy and exophagy and endophily and exophily will enhance our ability to develop and deploy vector-targeted interventions and monitoring techniques. Our laboratory-based analyses have revealed that crepuscular period photopreference differs between An. gambiae and An. stephensi. We employed qRT-PCR to assess crepuscular transcriptional expression patterns of long wavelength-, short wavelength-, and ultraviolet wavelength-sensing opsins (i.e., rhodopsin-class G-protein coupled receptors) in An. gambiae and in An. stephensi. Transcript levels do not exhibit consistent differences between species across diurnal cycles, indicating that differences in transcript abundances within this gene set are not correlated with these behavioral differences. Using developmentally staged and gender-specific RNAseq data sets in An. gambiae, we show that long wavelength-sensing opsins are expressed in two different patterns (one set expressed during larval stages, and one set expressed during adult stages), while short wavelength- and ultraviolet wavelength-sensing opsins exhibit increased expression during adult stages. Genomic organization of An. gambiae opsins suggests paralogous gene expansion of long wavelength-sensing opsins in comparison with An. stephensi. We speculate that this difference in gene number may contribute to variation between these species in photopreference behavior (e.g., visual sensitivity). PMID:26334802

Dose-response tests and semi-field evaluation of lethal and sub-lethal effects of slow release pyriproxyfen granules (Sumilarv®0.5G) for the control of the malaria vectors Anopheles gambiae sensu lato.

PubMed

Mbare, Oscar; Lindsay, Steven W; Fillinger, Ulrike

2013-03-14

Recently research has shown that larviciding can be an effective tool for integrated malaria vector control. Nevertheless, the uptake of this intervention has been hampered by the need to re-apply larvicides frequently. There is a need to explore persistent, environmentally friendly larvicides for malaria vector control to reduce intervention efforts and costs by reducing the frequency of application. In this study, the efficacy of a 0.5% pyriproxyfen granule (Surmilarv®0.5G, Sumitomo Chemicals) was assessed for the control of Anopheles gambiae sensu stricto and Anopheles arabiensis, the major malaria vectors in sub-Saharan Africa. Dose-response and standardized field tests were implemented following standard procedures of the World Health Organization's Pesticide Evaluation Scheme to determine: (i) the susceptibility of vectors to this formulation; (ii) the residual activity and appropriate retreatment schedule for field application; and, (iii) sub-lethal impacts on the number and viability of eggs laid by adults after exposure to Sumilarv®0.5G during larval development. Anopheles gambiae s.s. and An. arabiensis were highly susceptible to Sumilarv®0.5G. Estimated emergence inhibition (EI) values were very low and similar for both species. The minimum dosage that completely inhibited adult emergence was between 0.01-0.03 parts per million (ppm) active ingredient (ai). Compared to the untreated control, an application of 0.018 ppm ai prevented 85% (95% confidence interval (CI) 82%-88%) of adult emergence over six weeks under standardized field conditions. A fivefold increase in dosage of 0.09 ppm ai prevented 97% (95% CI 94%-98%) emergence. Significant sub-lethal effects were observed in the standardized field tests. Female An. gambiae s.s. that were exposed to 0.018 ppm ai as larvae laid 47% less eggs, and females exposed to 0.09 ppm ai laid 74% less eggs than females that were unexposed to the treatment. Furthermore, 77% of eggs laid by females exposed to 0

2La chromosomal inversion enhances thermal tolerance of Anopheles gambiae larvae.

PubMed

Rocca, Kyle A C; Gray, Emilie M; Costantini, Carlo; Besansky, Nora J

2009-07-02

The mosquito Anopheles gambiae is broadly distributed throughout sub-Saharan Africa and this contributes to making it the most efficient vector of malaria on the continent. The pervasiveness of this species is hypothesized to originate in local adaptations facilitated by inversion polymorphisms. One inversion, named 2La, is strongly associated with aridity clines in West and Central Africa: while 2La is fixed in arid savannas, the 2L+a arrangement is predominantly found in the rainforest. Ability to survive high temperature exposure is an essential component of aridity tolerance, particularly in immature stages that are restricted to shallow puddles. Toward deciphering the role of the 2La inversion in local adaptation, the present investigation focused on variation in larval and pupal thermo-tolerance in two populations dissimilar solely in 2La arrangement. A laboratory colony of A. gambiae that is polymorphic for 2La but standard for all other known inversions was used to create 2 homokaryotypic populations (2L+a and 2La). The survival of 4th instar larvae and pupae from both populations was then tested following exposure to thermal stress with and without prior heat hardening. Larvae responded identically to a 40 degrees C heat stress, with about 50% of larvae dying after 1.5-2 h and few larvae surviving a 3 h stress. When heat hardened prior to the thermal stress, thermo-tolerance of both larval populations increased, with 2La 24 h survival significantly exceeding that of 2L+a. Pupae were generally more thermo-tolerant than larvae, although 2La pupae were less so than 2L+a. Heat hardening had no positive effect on pupal thermo-tolerance. The increased thermo-tolerance observed in 2La larvae following heat hardening suggests higher responsiveness (i.e., thermal sensitivity) of the inverted karyotype. By responding more drastically to the heat shock, 2La larvae are better equipped to resist the potentially lethal temperatures that occur in arid habitats. The lower

Molecular Basis for Genetic Resistance of Anopheles gambiae to Plasmodium: Structural Analysis of TEP1 Susceptible and Resistant Alleles

PubMed Central

Logarajah, Shankar; Baxter, Richard H. G.

2012-01-01

Thioester-containing protein 1 (TEP1) is a central component in the innate immune response of Anopheles gambiae to Plasmodium infection. Two classes of TEP1 alleles, TEP1*S and TEP1*R, are found in both laboratory strains and wild isolates, related by a greater or lesser susceptibility, respectively to both P. berghei and P. falciparum infection. We report the crystal structure of the full-length TEP1*S1 allele which, while similar to the previously determined structure of full-length TEP1*R1, displays flexibility in the N-terminal fragment comprising domains MG1-MG6. Amino acid differences between TEP1*R1 and TEP1*S1 are localized to the TED-MG8 domain interface that protects the thioester bond from hydrolysis and structural changes are apparent at this interface. As a consequence cleaved TEP1*S1 (TEP1*S1cut) is significantly more susceptible to hydrolysis of its intramolecular thioester bond than TEP1*R1cut. TEP1*S1cut is stabilized in solution by the heterodimeric LRIM1/APL1C complex, which preserves the thioester bond within TEP1*S1cut. These results suggest a mechanism by which selective pressure on the TEP1 gene results in functional variation that may influence the vector competence of A. gambiae towards Plasmodium infection. PMID:23055931

Molecular basis for genetic resistance of Anopheles gambiae to Plasmodium: structural analysis of TEP1 susceptible and resistant alleles.

PubMed

Le, Binh V; Williams, Marni; Logarajah, Shankar; Baxter, Richard H G

2012-01-01

Thioester-containing protein 1 (TEP1) is a central component in the innate immune response of Anopheles gambiae to Plasmodium infection. Two classes of TEP1 alleles, TEP1*S and TEP1*R, are found in both laboratory strains and wild isolates, related by a greater or lesser susceptibility, respectively to both P. berghei and P. falciparum infection. We report the crystal structure of the full-length TEP1*S1 allele which, while similar to the previously determined structure of full-length TEP1*R1, displays flexibility in the N-terminal fragment comprising domains MG1-MG6. Amino acid differences between TEP1*R1 and TEP1*S1 are localized to the TED-MG8 domain interface that protects the thioester bond from hydrolysis and structural changes are apparent at this interface. As a consequence cleaved TEP1*S1 (TEP1*S1(cut)) is significantly more susceptible to hydrolysis of its intramolecular thioester bond than TEP1*R1(cut). TEP1*S1(cut) is stabilized in solution by the heterodimeric LRIM1/APL1C complex, which preserves the thioester bond within TEP1*S1(cut). These results suggest a mechanism by which selective pressure on the TEP1 gene results in functional variation that may influence the vector competence of A. gambiae towards Plasmodium infection.

Molecular and functional characterization of Anopheles gambiae inward rectifier potassium (Kir1) channels: a novel role in egg production.

PubMed

Raphemot, Rene; Estévez-Lao, Tania Y; Rouhier, Matthew F; Piermarini, Peter M; Denton, Jerod S; Hillyer, Julián F

2014-08-01

Inward rectifier potassium (Kir) channels play essential roles in regulating diverse physiological processes. Although Kir channels are encoded in mosquito genomes, their functions remain largely unknown. In this study, we identified the members of the Anopheles gambiae Kir gene family and began to investigate their function. Notably, we sequenced the A. gambiae Kir1 (AgKir1) gene and showed that it encodes all the canonical features of a Kir channel: an ion pore that is composed of a pore helix and a selectivity filter, two transmembrane domains that flank the ion pore, and the so-called G-loop. Heterologous expression of AgKir1 in Xenopus oocytes revealed that this gene encodes a functional, barium-sensitive Kir channel. Quantitative RT-PCR experiments then showed that relative AgKir1 mRNA levels are highest in the pupal stage, and that AgKir1 mRNA is enriched in the adult ovaries. Gene silencing of AgKir1 by RNA interference did not affect the survival of female mosquitoes following a blood meal, but decreased their egg output. These data provide evidence for a new role of Kir channels in mosquito fecundity, and further validates them as promising molecular targets for the development of a new class of mosquitocides to be used in vector control. Copyright © 2014 Elsevier Ltd. All rights reserved.

Analysis of expression in the Anopheles gambiae developing testes reveals rapidly evolving lineage-specific genes in mosquitoes.

PubMed

Krzywinska, Elzbieta; Krzywinski, Jaroslaw

2009-07-06

Male mosquitoes do not feed on blood and are not involved in delivery of pathogens to humans. Consequently, they are seldom the subjects of research, which results in a very poor understanding of their biology. To gain insights into male developmental processes we sought to identify genes transcribed exclusively in the reproductive tissues of male Anopheles gambiae pupae. Using a cDNA subtraction strategy, five male-specifically or highly male-biased expressed genes were isolated, four of which remain unannotated in the An. gambiae genome. Spatial and temporal expression patterns suggest that each of these genes is involved in the mid-late stages of spermatogenesis. Their sequences are rapidly evolving; however, two genes possess clear homologs in a wide range of taxa and one of these probably acts in a sperm motility control mechanism conserved in many organisms, including humans. The other three genes have no match to sequences from non-mosquito taxa, thus can be regarded as orphans. RNA in situ hybridization demonstrated that one of the orphans is transcribed in spermatids, which suggests its involvement in sperm maturation. Two other orphans have unknown functions. Expression analysis of orthologs of all five genes indicated that male-biased transcription was not conserved in the majority of cases in Aedes and Culex. Discovery of testis-expressed orphan genes in mosquitoes opens new prospects for the development of innovative control methods. The orphan encoded proteins may represent unique targets of selective anti-mosquito sterilizing agents that will not affect non-target organisms.

Spatial and temporal heterogeneity of Anopheles mosquitoes and Plasmodium falciparum transmission along the Kenyan coast.

PubMed

Mbogo, Charles M; Mwangangi, Joseph M; Nzovu, Joseph; Gu, Weidong; Yan, Guiyan; Gunter, James T; Swalm, Chris; Keating, Joseph; Regens, James L; Shililu, Josephat I; Githure, John I; Beier, John C

2003-06-01

The seasonal dynamics and spatial distributions of Anopheles mosquitoes and Plasmodium falciparum parasites were studied for one year at 30 villages in Malindi, Kilifi, and Kwale Districts along the coast of Kenya. Anopheline mosquitoes were sampled inside houses at each site once every two months and malaria parasite prevalence in local school children was determined at the end of the entomologic survey. A total of 5,476 Anopheles gambiae s.l. and 3,461 An. funestus were collected. Species in the An. gambiae complex, identified by a polymerase chain reaction, included 81.9% An. gambiae s.s., 12.8% An. arabiensis, and 5.3% An. merus. Anopheles gambiae s.s. contributed most to the transmission of P. falciparum along the coast as a whole, while An. funestus accounted for more than 50% of all transmission in Kwale District. Large spatial heterogeneity of transmission intensity (< 1 up to 120 infective bites per person per year) resulted in correspondingly large and significantly related variations in parasite prevalence (range = 38-83%). Thirty-two percent of the sites (7 of 22 sites) with malaria prevalences ranging from 38% to 70% had annual entomologic inoculation rates (EIR) less than five infective bites per person per year. Anopheles gambiae s.l. and An. funestus densities in Kwale were not significantly influenced by rainfall. However, both were positively correlated with rainfall one and three months previously in Malindi and Kilifi Districts, respectively. These unexpected variations in the relationship between mosquito populations and rainfall suggest environmental heterogeneity in the predominant aquatic habitats in each district. One important conclusion is that the highly non-linear relationship between EIRs and prevalence indicates that the consistent pattern of high prevalence might be governed by substantial variation in transmission intensity measured by entomologic surveys. The field-based estimate of entomologic parameters on a district level does

Diversity in breeding sites and distribution of Anopheles mosquitoes in selected urban areas of southern Ghana.

PubMed

Mattah, Precious A Dzorgbe; Futagbi, Godfred; Amekudzi, Leonard K; Mattah, Memuna M; de Souza, Dziedzorm K; Kartey-Attipoe, Worlasi D; Bimi, Langbong; Wilson, Michael D

2017-01-13

Anopheles vectors of malaria are supposedly less common in urban areas as a result of pollution, but there is increasing evidence of their adaptation to organically polluted water bodies. This study characterized the breeding habitats of Anopheles mosquitoes in the two major urban areas in southern Ghana; Accra (AMA) and Sekondi-Takoradi (STMA) Metropolitan Areas, during dry and wet seasons. Anopheles mosquito larvae were sampled using standard dipping methods to determine larval densities. The origin, nature and stability of 21 randomly selected sites were observed and recorded. Mosquito larvae were reared to adults and Anopheles species identified by both morphological and molecular means. Sixty-six percent of Anopheles habitats were permanent and 34% temporal, and 74.5% man-made while 25.5% were natural. Puddles and urban farm sites accounted for over 51% of all Anopheles mosquitoes sampled. The mean larval densities among the habitat types was highest of 13.7/dip for puddles and lowest of 2.3/dip for stream/river, and the variation between densities were significant (P = 0.002). The mean larval densities were significantly higher in the wet season than in the dry season for the two study areas combined (P = 0.0191) and AMA (P = 0.0228). Over 99% of the 5,802 morphologically identified Anopheles species were An. gambiae (s.l.) of which more than 99% of the studied 898 were An. coluzzii (62%) and An. gambiae (s.s.) (34%). Urban farms, puddles, swamps and ditches/ dugouts accounted for approximately 70% of all An. coluzzii identified. Conversely, drains, construction sites, streams/rivers and "others" contributed 80% of all An. gambiae (s.s.) sampled. The wet season had significantly higher proportion of Anopheles larvae compared to the dry season (Z = 8.3683, P < 0.0001). Also, the proportion of Anopheles mosquitoes produced by permanent breeding sites was 61.3% and that of temporary sites was 38.7%. Taken together, the data suggest that man

Spatial distribution and habitat characterisation of Anopheles larvae along the Kenyan coast.

PubMed

Mwangangi, Joseph M; Mbogo, Charles M; Muturi, Ephantus J; Nzovu, Joseph G; Githure, John I; Yan, Guiyun; Minakawa, Noboru; Novak, Robert; Beier, John C

2007-03-01

A study was conducted to characterise larval habitats and to determine spatial heterogeneity of the Anopheles mosquito larvae. The study was conducted from May to June 1999 in nine villages along the Kenyan coast. Aquatic habitats were sampled by use of standard dipping technique. The habitats were characterised based on size, pH, distance to the nearest house, coverage of canopy, surface debris, algae and emergent plants, turbidity, substrate, and habitat type. A total of 110 aquatic habitats like stream pools (n=10); puddles (n=65); tire tracks (n=5); ponds (n=5) and swamps (n=25) were sampled in nine villages located in three districts of the Kenyan coast. A total of 7,263 Anopheles mosquito larvae were collected, 63.9% were early instars and 36.1% were late instars. Morphological identification of the III and IV instar larvae by use of microscopy yielded 90.66% (n=2377) Anopheles gambiae Complex, 0.88% (n=23) An. funestus, An. coustani 7.63% (n=200), An. rivulorum 0.42% (n=11), An. pharoensis 0.19% (n=5), An. swahilicus 0.08% (n=2), An. wilsoni 0.04% (n=1) and 0.11% (n=3) were unidentified. A subset of the An. gambiae Complex larvae identified morphologically, was further analysed using rDNA-PCR technique resulting in 68.22% (n=1290) An. gambiae s.s., 7.93% (n=150) An. arabiensis and 23.85% (n=451) An. merus. Multiple logistic regression model showed that emergent plants (p = 0.019), and floating debris (p = 0.038) were the best predictors of An. gambiae larval abundance in these habitats. Habitat type, floating debris and emergent plants were found to be the key factors determining the presence of Anopheles larvae in the habitats. For effective larval control, the type of habitat should be considered and most productive habitat type be given a priority in the mosquito abatement programme.

Status of pyrethroid resistance in Anopheles gambiae s. s. M form prior to the scaling up of Long Lasting Insecticidal Nets (LLINs) in Adzopé, Eastern Côte d’Ivoire

PubMed Central

2012-01-01

Background The growing development of pyrethroid resistance constitutes a serious threat to malaria control programmes and if measures are not taken in time, resistance may compromise control efforts in the foreseeable future. Prior to Long Lasting Insecticidal Nets (LLINs) distribution in Eastern Cote d’Ivoire, we conducted bioassays to inform the National Malaria Control Programme of the resistance status of the main malaria vector, Anopheles gambiae s. s. and the need for close surveillance of resistance. Methods Larvae of An. gambiae s. s. were collected in two areas of Adzopé (Port-Bouët and Tsassodji) and reared to adults. WHO susceptibility tests with impregnated filter papers were carried out to detect resistance to three pyrethroids commonly used to develop LLINs: permethrin 1%, deltamethrin 0.05% and lambda-cyhalothrin 0.05%. Molecular assays were conducted to detect M and S forms and the L1014F kdr allele in individual mosquitoes. Results Resistance, at various degrees was detected in both areas of Adzopé. Overall, populations of An. gambiae at both sites surveyed showed equivalent frequency of the L1014F kdr allele (0.67) but for all tested pyrethroids, there were significantly higher survival rates for mosquitoes from Tsassodji (32–58%) than those from Port-Bouët (3–32%) (p < 0.001), indicating the implication of resistance mechanisms other than kdr alone. During the survey period (May–June) in this forested area of Côte d’Ivoire, An. gambiae s. s. found were exclusively of the M form and were apparently selected for pyrethroid resistance through agricultural and household usage of insecticides. Conclusion Prior to LLINs scaling up in Eastern Côte d’Ivoire, resistance was largely present at various levels in An. gambiae. Underlying mechanisms included the high frequency of the L1014F kdr mutation and other unidentified components, probably metabolic detoxifiers. Their impact on the efficacy of the planned strategy (LLINs) in the

Long-term trends in Anopheles gambiae insecticide resistance in Côte d'Ivoire.

PubMed

Edi, Constant A V; Koudou, Benjamin G; Bellai, Louise; Adja, Akre M; Chouaibou, Mouhamadou; Bonfoh, Bassirou; Barry, Sarah J E; Johnson, Paul C D; Müller, Pie; Dongus, Stefan; N'Goran, Eliezer K; Ranson, Hilary; Weetman, David

2014-11-28

Malaria control is heavily dependent on the use of insecticides that target adult mosquito vectors via insecticide treated nets (ITNs) or indoor residual spraying (IRS). Four classes of insecticide are approved for IRS but only pyrethroids are available for ITNs. The rapid rise in insecticide resistance in African malaria vectors has raised alarms about the sustainability of existing malaria control activities. This problem might be particularly acute in Côte d'Ivoire where resistance to all four insecticide classes has recently been recorded. Here we investigate temporal trends in insecticide resistance across the ecological zones of Côte d'Ivoire to determine whether apparent pan-African patterns of increasing resistance are detectable and consistent across insecticides and areas. We combined data on insecticide resistance from a literature review, and bioassays conducted on field-caught Anopheles gambiae mosquitoes for the four WHO-approved insecticide classes for ITN/IRS. The data were then mapped using Geographical Information Systems (GIS) and the IR mapper tool to provide spatial and temporal distribution data on insecticide resistance in An. gambiae sensu lato from Côte d'Ivoire between 1993 and 2014. Bioassay mortality decreased over time for all insecticide classes, though with significant spatiotemporal variation, such that stronger declines were observed in the southern ecological zone for DDT and pyrethroids than in the central zone, but with an apparently opposite effect for the carbamate and organophosphate. Variation in relative abundance of the molecular forms, coupled with dramatic increase in kdr 1014F frequency in M forms (An. coluzzii) seems likely to be a contributory factor to these patterns. Although records of resistance across insecticide classes have become more common, the number of classes tested in studies has also increased, precluding a conclusion that multiple resistance has also increased. Our analyses attempted synthesis of 22�

Larvicidal potential of some plants from West Africa against Culex quinquefasciatus (Say) and Anopheles gambiae Giles (Diptera: Culicidae).

PubMed

Azokou, Alain; Koné, Mamidou W; Koudou, Benjamin G; Tra Bi, Honora F

2013-01-01

Mosquitoes increased resistance to insecticides, and environmental concerns about the use of insecticides, pose a major challenge in the search for new molecules to deplete and incapacitate mosquito populations. Plants are the valuable source as practices consisting in exploiting plant materials as repellents, and are still in wide use throughout developing countries. The aim of the present study was to screen plants from Cτte d'Ivoire for larvicidal activity against mosquitoes. Resistant and sensitive larvae (III and IV instar) of Anopheles gambiae and Culex quinquefasciatus were exposed to crude ethanol extracts (90%) of 45 plants and viability observed after 30 min, 6, 12 and 24 h postincubation. After partition of active extracts, each fraction (hexane and chloroform washed with NaCl 1%, tannins and aqueous) was tested using the same protocol at various concentrations (1000- 31.2 ppm). Of 49 extracts tested, 7 exhibited high potential (LC50 = 80 to 370 ppm) against resistant and sensitive III and IV instar larvae of An. gambiae and Cx. quinquefasciatus. These extracts were from Cissus populnea, Cochlospermum planchonii, Heliotropium indicum, Phyllanthus amarus, Vitex grandifolia and Alchornea cordifolia. However, three most active plant species (LC50 = 80- 180 ppm) were Cs. populnea, Cm. planchonii and P. amarus Their hexane and chloroform fractions showed high larvicidal activity. This study demonstrated that plants from Cτte d'Ivoire have a real potential for malaria, yellow fever, filarial and dengue vector control. Those could be used as sources or provide lead compounds for the development of safe plant-based biocides.

Divergent transcriptional response to thermal stress by Anopheles gambiae larvae carrying alternative arrangements of inversion 2La.

PubMed

Cassone, Bryan J; Molloy, Matthew J; Cheng, Changde; Tan, John C; Hahn, Matthew W; Besansky, Nora J

2011-06-01

The African malaria mosquito Anopheles gambiae is polymorphic for chromosomal inversion 2La, whose frequency strongly correlates with degree of aridity across environmental gradients. Recent physiological studies have associated 2La with resistance to desiccation in adults and thermal stress in larvae, consistent with its proposed role in aridity tolerance. However, the genetic basis of these traits remains unknown. To identify genes that could be involved in the differential response to thermal stress, we compared global gene expression profiles of heat-hardened 2La or 2L+(a) larvae at three time points, for up to eight hours following exposure to the heat stress. Treatment and control time series, replicated four times, revealed a common and massive induction of a core set of heat-shock genes regardless of 2La orientation. However, clear differences between the 2La and 2L+(a) arrangements emerged at the earliest (0.25 h) time point, in the intensity and nature of the stress response. Overall, 2La was associated with the more aggressive response: larger numbers of genes were heat responsive and up-regulated. Transcriptionally induced genes were enriched for functions related to ubiquitin-proteasomal degradation, chaperoning and energy metabolism. The more muted transcriptional response of 2L+(a) was largely repressive, including genes involved in proteolysis and energy metabolism. These results may help explain the maintenance of the 2La inversion polymorphism in An. gambiae, as the survival benefits offered by high thermal sensitivity in harsh climates could be offset by the metabolic costs of such a drastic response in more equable climates. © 2011 Blackwell Publishing Ltd.

Rickettsia Species in African Anopheles Mosquitoes

PubMed Central

Socolovschi, Cristina; Pages, Frédéric; Ndiath, Mamadou O.; Ratmanov, Pavel; Raoult, Didier

2012-01-01

Background There is higher rate of R. felis infection among febrile patients than in healthy people in Sub-Saharan Africa, predominantly in the rainy season. Mosquitoes possess a high vectorial capacity and, because of their abundance and aggressiveness, likely play a role in rickettsial epidemiology. Methodology/Principal Findings Quantitative and traditional PCR assays specific for Rickettsia genes detected rickettsial DNA in 13 of 848 (1.5%) Anopheles mosquitoes collected from Côte d’Ivoire, Gabon, and Senegal. R. felis was detected in one An. gambiae molecular form S mosquito collected from Kahin, Côte d’Ivoire (1/77, 1.3%). Additionally, a new Rickettsia genotype was detected in five An. gambiae molecular form S mosquitoes collected from Côte d’Ivoire (5/77, 6.5%) and one mosquito from Libreville, Gabon (1/88, 1.1%), as well as six An. melas (6/67, 9%) mosquitoes collected from Port Gentil, Gabon. A sequence analysis of the gltA, ompB, ompA and sca4 genes indicated that this new Rickettsia sp. is closely related to R. felis. No rickettsial DNA was detected from An. funestus, An. arabiensis, or An. gambiae molecular form M mosquitoes. Additionally, a BLAST analysis of the gltA sequence from the new Rickettsia sp. resulted in a 99.71% sequence similarity to a species (JQ674485) previously detected in a blood sample of a Senegalese patient with a fever from the Bandafassi village, Kedougou region. Conclusion R. felis was detected for the first time in An. gambiae molecular form S, which represents the major African malaria vector. The discovery of R. felis, as well as a new Rickettsia species, in mosquitoes raises new issues with respect to African rickettsial epidemiology that need to be investigated, such as bacterial isolation, the degree of the vectorial capacity of mosquitoes, the animal reservoirs, and human pathogenicity. PMID:23118963

Reflections on the Anopheles gambiae genome sequence, transgenic mosquitoes and the prospect for controlling malaria and other vector borne diseases.

PubMed

Tabachnick, Walter J

2003-09-01

The completion of the Anopheles gambiae Giles genome sequencing project is a milestone toward developing more effective strategies in reducing the impact of malaria and other vector borne diseases. The successes in developing transgenic approaches using mosquitoes have provided another essential new tool for further progress in basic vector genetics and the goal of disease control. The use of transgenic approaches to develop refractory mosquitoes is also possible. The ability to use genome sequence to identify genes, and transgenic approaches to construct refractory mosquitoes, has provided the opportunity that with the future development of an appropriate genetic drive system, refractory transgenes can be released into vector populations leading to nontransmitting mosquitoes. An. gambiae populations incapable of transmitting malaria. This compelling strategy will be very difficult to achieve and will require a broad substantial research program for success. The fundamental information that is required on genome structure, gene function and environmental effects on genetic expression are largely unknown. The ability to predict gene effects on phenotype is rudimentary, particularly in natural populations. As a result, the release of a refractory transgene into natural mosquito populations is imprecise and there is little ability to predict unintended consequences. The new genetic tools at hand provide opportunities to address an array of important issues, many of which can have immediate impact on the effectiveness of a host of strategies to control vector borne disease. Transgenic release approaches represent only one strategy that should be pursued. A balanced research program is required.

Diel timing and frequency of sugar feeding in the mosquito Anopheles gambiae, depending on sex, gonotrophic state and resource availability.

PubMed

Gary, R E; Foster, W A

2006-09-01

Little is known about the sugar-feeding behaviour of equatorial Africa's principal vector of malaria, Anopheles gambiae Giles (Diptera: Culicidae). It is suspected to feed on plant sugar infrequently, but possibly the timing depends on environmental circumstances, and males may differ markedly from females. These points of uncertainty were clarified in the laboratory, by monitoring both diel and longterm sugar-feeding activity in both sexes. Males fed on sugar in a nocturnal diel rhythm closely approximating non-specific flight activity. Female diel sugar-feeding patterns resembled published rhythms and cycles of host seeking. Males sugar fed nightly at an average frequency of about twice per night, sustained over 17 days. This was substantially higher than the sugar-feeding frequency of females that were allowed both blood and oviposition sites every night: they averaged about one sugar feed in every 4 nights. These females fed on sugar between gonotrophic cycles, after eggs were mature but before the next bloodmeal. They did not sugar feed during the 2 days after blood feeding, while blood was being digested and the eggs developed. A slight delay in the availability of either the oviposition site or blood led to an increase in female sugar-feeding frequency: they averaged more than once per night until the delayed resource was made available. These observations support the conclusion that sugar feeding is a normal part of the biology of both sexes of An. gambiae.

Anthropogenic habitat disturbance and ecological divergence between incipient species of the malaria mosquito Anopheles gambiae.

PubMed

Kamdem, Colince; Tene Fossog, Billy; Simard, Frédéric; Etouna, Joachim; Ndo, Cyrille; Kengne, Pierre; Boussès, Philippe; Etoa, François-Xavier; Awono-Ambene, Parfait; Fontenille, Didier; Antonio-Nkondjio, Christophe; Besansky, Nora J; Costantini, Carlo

2012-01-01

Anthropogenic habitat disturbance is a prime cause in the current trend of the Earth's reduction in biodiversity. Here we show that the human footprint on the Central African rainforest, which is resulting in deforestation and growth of densely populated urban agglomerates, is associated to ecological divergence and cryptic speciation leading to adaptive radiation within the major malaria mosquito Anopheles gambiae. In southern Cameroon, the frequency of two molecular forms--M and S--among which reproductive isolation is strong but still incomplete, was correlated to an index of urbanisation extracted from remotely sensed data, expressed as the proportion of built-up surface in each sampling unit. The two forms markedly segregated along an urbanisation gradient forming a bimodal cline of ∼6-km width: the S form was exclusive to the rural habitat, whereas only the M form was present in the core of densely urbanised settings, co-occurring at times in the same polluted larval habitats of the southern house mosquito Culex quinquefasciatus--a species association that was not historically recorded before. Our results indicate that when humans create novel habitats and ecological heterogeneities, they can provide evolutionary opportunities for rapid adaptive niche shifts associated with lineage divergence, whose consequences upon malaria transmission might be significant.

Insecticide resistance profile of Anopheles gambiae from a phase II field station in Cové, southern Benin: implications for the evaluation of novel vector control products.

PubMed

Ngufor, Corine; N'Guessan, Raphael; Fagbohoun, Josias; Subramaniam, Krishanthi; Odjo, Abibatou; Fongnikin, Augustin; Akogbeto, Martin; Weetman, David; Rowland, Mark

2015-11-18

Novel indoor residual spraying (IRS) and long-lasting insecticidal net (LLIN) products aimed at improving the control of pyrethroid-resistant malaria vectors have to be evaluated in Phase II semi-field experimental studies against highly pyrethroid-resistant mosquitoes. To better understand their performance it is necessary to fully characterize the species composition, resistance status and resistance mechanisms of the vector populations in the experimental hut sites. Bioassays were performed to assess phenotypic insecticide resistance in the malaria vector population at a newly constructed experimental hut site in Cové, a rice growing area in southern Benin, being used for WHOPES Phase II evaluation of newly developed LLIN and IRS products. The efficacy of standard WHOPES-approved pyrethroid LLIN and IRS products was also assessed in the experimental huts. Diagnostic genotyping techniques and microarray studies were performed to investigate the genetic basis of pyrethroid resistance in the Cové Anopheles gambiae population. The vector population at the Cové experimental hut site consisted of a mixture of Anopheles coluzzii and An. gambiae s.s. with the latter occurring at lower frequencies (23 %) and only in samples collected in the dry season. There was a high prevalence of resistance to pyrethroids and DDT (>90 % bioassay survival) with pyrethroid resistance intensity reaching 200-fold compared to the laboratory susceptible An. gambiae Kisumu strain. Standard WHOPES-approved pyrethroid IRS and LLIN products were ineffective in the experimental huts against this vector population (8-29 % mortality). The L1014F allele frequency was 89 %. CYP6P3, a cytochrome P450 validated as an efficient metabolizer of pyrethroids, was over-expressed. Characterizing pyrethroid resistance at Phase II field sites is crucial to the accurate interpretation of the performance of novel vector control products. The strong levels of pyrethroid resistance at the Cové experimental hut

Modeling the role of environmental variables on the population dynamics of the malaria vector Anopheles gambiae sensu stricto

PubMed Central

2012-01-01

Background The impact of weather and climate on malaria transmission has attracted considerable attention in recent years, yet uncertainties around future disease trends under climate change remain. Mathematical models provide powerful tools for addressing such questions and understanding the implications for interventions and eradication strategies, but these require realistic modeling of the vector population dynamics and its response to environmental variables. Methods Published and unpublished field and experimental data are used to develop new formulations for modeling the relationships between key aspects of vector ecology and environmental variables. These relationships are integrated within a validated deterministic model of Anopheles gambiae s.s. population dynamics to provide a valuable tool for understanding vector response to biotic and abiotic variables. Results A novel, parsimonious framework for assessing the effects of rainfall, cloudiness, wind speed, desiccation, temperature, relative humidity and density-dependence on vector abundance is developed, allowing ease of construction, analysis, and integration into malaria transmission models. Model validation shows good agreement with longitudinal vector abundance data from Tanzania, suggesting that recent malaria reductions in certain areas of Africa could be due to changing environmental conditions affecting vector populations. Conclusions Mathematical models provide a powerful, explanatory means of understanding the role of environmental variables on mosquito populations and hence for predicting future malaria transmission under global change. The framework developed provides a valuable advance in this respect, but also highlights key research gaps that need to be resolved if we are to better understand future malaria risk in vulnerable communities. PMID:22877154

Evaluation of two counterflow traps for testing behaviour-mediating compounds for the malaria vector Anopheles gambiae s.s. under semi-field conditions in Tanzania

PubMed Central

Schmied, Wolfgang H; Takken, Willem; Killeen, Gerry F; Knols, Bart GJ; Smallegange, Renate C

2008-01-01

Background Evaluation of mosquito responses towards different trap-bait combinations in field trials is a time-consuming process that can be shortened by experiments in contained semi-field systems. Possible use of the BG Sentinel (BGS) trap to sample Anopheles gambiae s.s. was evaluated. The efficiency of this trap was compared with that of the Mosquito Magnet-X (MM-X) trap, when baited with foot odour alone or combinations of foot odour with carbon dioxide (CO2) or lemongrass as behaviour-modifying cues. Methods Female An. gambiae s.s. were released in an experimental flight arena that was placed in a semi-field system and left overnight. Catch rates for the MM-X and BGS traps were recorded. Data were analysed by fitting a generalized linear model to the (n+1) transformed catches. Results Both types of traps successfully captured mosquitoes with all odour cues used. When the BGS trap was tested against the MM-X trap in a choice assay with foot odour as bait, the BGS trap caught about three times as many mosquitoes as the MM-X trap (P = 0.002). Adding CO2 (500 ml/min) to foot odour increased the number of mosquitoes caught by 268% for the MM-X (P < 0.001) and 34% (P = 0.051) for the BGS trap, compared to foot odour alone. When lemongrass leaves were added to foot odour, mosquito catches were reduced by 39% (BGS, P < 0.001) and 38% (MM-X, P = 0.353), respectively. Conclusion The BGS trap shows high potential for field trials due to its simple construction and high catch rate when baited with human foot odour only. However, for rapid screening of different baits in a contained semi-field system, the superior discriminatory power of the MM-X trap is advantageous. PMID:18980669

Clustering of host-seeking activity of Anopheles gambiae mosquitoes at the top surface of a human-baited bed net

PubMed Central

2013-01-01

Background Knowledge of the interactions between mosquitoes and humans, and how vector control interventions affect them, is sparse. A study exploring host-seeking behaviour at a human-occupied bed net, a key event in such interactions, is reported here. Methods Host-seeking female Anopheles gambiae activity was studied using a human-baited ‘sticky-net’ (a bed net without insecticide, coated with non-setting adhesive) to trap mosquitoes. The numbers and distribution of mosquitoes captured on each surface of the bed net were recorded and analysed using non-parametric statistical methods and random effects regression analysis. To confirm sticky-net reliability, the experiment was repeated using a pitched sticky-net (tilted sides converging at apex, i.e., neither horizontal nor vertical). The capture efficiency of horizontal and vertical sticky surfaces were compared, and the potential repellency of the adhesive was investigated. Results In a semi-field experiment, more mosquitoes were caught on the top (74-87%) than on the sides of the net (p < 0.001). In laboratory experiments, more mosquitoes were caught on the top than on the sides in human-baited tests (p < 0.001), significantly different to unbaited controls (p < 0.001) where most mosquitoes were on the sides (p = 0.047). In both experiments, approximately 70% of mosquitoes captured on the top surface were clustered within a 90 × 90 cm (or lesser) area directly above the head and chest (p < 0.001). In pitched net tests, similar clustering occurred over the sleeper’s head and chest in baited tests only (p < 0.001). Capture rates at horizontal and vertical surfaces were not significantly different and the sticky-net was not repellent. Conclusion This study demonstrated that An. gambiae activity occurs predominantly within a limited area of the top surface of bed nets. The results provide support for the two-in-one bed net design for managing pyrethroid-resistant vector populations

Effects of larval growth condition and water availability on desiccation resistance and its physiological basis in adult Anopheles gambiae sensu stricto

PubMed Central

2010-01-01

Background Natural populations of the malaria mosquito Anopheles gambiae s.s. are exposed to large seasonal and daily fluctuations in relative humidity and temperature, which makes coping with drought a crucial aspect of their ecology. Methods To better understand natural variation in desiccation resistance in this species, the effects of variation in larval food availability and access to water as an adult on subsequent phenotypic quality and desiccation resistance of adult females of the Mopti chromosomal form were tested experimentally. Results It was found that, under normal conditions, larval food availability and adult access to water had only small direct effects on female wet mass, dry mass, and water, glycogen and body lipid contents corrected for body size. In contrast, when females subsequently faced a strong desiccation challenge, larval food availability and adult access to water had strong carry-over effects on most measured physiological and metabolic parameters, and affected female survival. Glycogen and water content were the most used physiological reserves in relative terms, but their usage significantly depended on female phenotypic quality. Adult access to water significantly influenced the use of water and body lipid reserves, which subsequently affected desiccation resistance. Conclusions These results demonstrate the importance of growth conditions and water availability on adult physiological status and subsequent resistance to desiccation. PMID:20691104

Effects of larvicidal and larval nutritional stresses on Anopheles gambiae development, survival and competence for Plasmodium falciparum.

PubMed

Vantaux, Amélie; Ouattarra, Issiaka; Lefèvre, Thierry; Dabiré, Kounbobr Roch

2016-04-23

Many studies have shown that the environment in which larvae develop can influence adult characteristics with consequences for the transmission of pathogens. We investigated how two environmental stresses (larviciding and nutritional stress) interact to affect Anopheles gambiae (previously An. gambiae S molecular form) life history traits and its susceptibility for field isolates of its natural malaria agent Plasmodium falciparum. Larvae were reared in the presence or not of a sub-lethal concentration of larvicide and under a high and low food regimen. Development time, individual size, adult survival and competence for P. falciparum were assessed. Individuals under low food regimen took more time to develop, had a lower development success and were smaller while there was no main effect of larvicide exposure on these traits. However, larvicide exposure impacted individual size in interaction with nutritional stress. Female survival was affected by the interaction between gametocytemia, parasite exposure and larval diet, as well as the interaction between gametocytemia, parasite exposure and larvicidal stress, and the interaction between gametocytemia, larvicidal exposure and larval diet. Among the 951 females dissected 7 days post-infection, 559 (58.78%) harboured parasites. Parasite prevalence was significantly affected by the interaction between larvicidal stress and larval diet. Indeed, females under low food regimen had a higher prevalence than females under high food regimen and this difference was greater under larvicidal stress. The two stresses did not impact parasite intensity. We found that larval nutritional and larvicidal stresses affect mosquito life history traits in complex ways, which could greatly affect P. falciparum transmission. Further studies combining field-based trials on larvicide use and mosquito experimental infections would give a more accurate understanding of the effects of this vector control tool on malaria transmission.

When intensity of deltamethrin resistance in Anopheles gambiae s.l. leads to loss of Long Lasting Insecticidal Nets bio-efficacy: a case study in north Cameroon.

PubMed

Etang, Josiane; Pennetier, Cédric; Piameu, Michael; Bouraima, Aziz; Chandre, Fabrice; Awono-Ambene, Parfait; Marc, Coosemans; Corbel, Vincent

2016-03-08

In Cameroon, insecticide resistance in Anopheles (An.) gambiae s.l. has been reported in several foci, prompting further investigations on associated patterns of Long-Lasting Insecticidal Nets (LLINs) bio-efficacy. The current study, conducted from June to August 2011, explored the intensity of deltamethrin resistance in An. gambiae s.l. from Pitoa and its impact on the residual bio-efficacy of LifeNet, a LLIN with deltamethrin incorporated into polypropylene nets (PND). Two-four days old females An. gambiae s.l. reared from larval collections in Pitoa were tested for susceptibility to DDT, permethrin and deltamethrin, using standard World Health Organization (WHO) tube assays. Intensity of deltamethrin resistance was explored using WHO tube assays, but across six working concentrations from 0.001 % to 0.5 %. Bio-efficacy of unwashed and washed PND was assessed using WHO cone test. Species identification and kdr 1014 genotyping were performed on mosquito samples that were not exposed to insecticides, using PCR-RFLP and HOLA methods respectively. The Kisumu reference susceptible strain of An. gambiae s.s. was used for comparisons. A total of 1895 An. gambiae s.l. specimens from Pitoa were used for resistance and PND bio-efficacy testing. This mosquito population was resistant to DDT, permethrin and deltamethrin, with 18-40 min knockdown times for 50 % of tested mosquitoes and 59-77 % mortality. Deltamethrin Resistance Ratio compared with the Kisumu strain was estimated at ≥500 fold. LifeNets were effective against the susceptible Kisumu (100 % knockdown (KD60min) and mortality) and the resistant Pitoa samples (95 % KD60min, 83-95 % mortality). However, the bio-efficacy gradually dropped against the Pitoa samples when nets were washed (X (2) = 35.887, df = 8, p < 0.001), and fell under the WHO efficacy threshold (80 % mortality and/or 95 % KD60min) between 10 and 15 washes. The Pitoa samples were composed of three sibling species: An. arabiensis (132

Decline in frequency of the 2La chromosomal inversion in Anopheles gambiae (s.s.) in Western Kenya: correlation with increase in ownership of insecticide-treated bed nets.

PubMed

Matoke-Muhia, Damaris; Gimnig, John E; Kamau, Luna; Shililu, Josephat; Bayoh, M Nabie; Walker, Edward D

2016-06-10

The 2La chromosomal inversion, a genetic polymorphism in An. gambiae (sensu stricto) (s.s.), is associated with adaptation to microclimatic differences in humidity and desiccation resistance and mosquito behaviors. Ownership of insecticide-treated bed nets (ITNs) for malaria control has increased markedly in western Kenya in the last 20 years. An increase in the frequency of ITNs indoors could select against house entering or indoor resting of Anopheles mosquitoes. Thus, the frequency of the 2La inversion is postulated to change in An. gambiae (s.s.) with the increase of ITN ownership over time. Anopheles gambiae mosquitoes were sampled between 1994 and 2011 using pyrethrum knockdown, bednet traps and human landing catches (HLC) from Asembo and Seme, western Kenya. The 2La inversion was detected by a PCR assay with primers designed for proximal breakpoints of the 2La/a and 2L+(a)/+(a) chromosomal conformation. Mosquitoes were tested for malaria parasite infection by sporozoite ELISA. The frequency of the 2La chromosomal inversion declined from 100 % of all chromosomes in 1994 to 17 % in 2005 and remained low through 2011 (21 %). ITN ownership increased from 0 to > 90 % of houses in the study area during this interval. The decline in the frequency of the 2La chromosomal inversion was significantly, negatively correlated with year (r = -0.93) and with increase in ITN ownership (r = -0.96). The frequency of the homo- and heterokaryotypes departed significantly from Hardy-Weinberg equilibrium, suggesting that 2La/a karyotype was under selection, earlier in its favor and later, against it. Precipitation and maximum monthly temperature did not vary over time, therefore there was no trend in climate that could account for the decline. There was no significant difference in frequency of the 2La inversion in An. gambiae (s.s.) females sampled indoors or outdoors in HCL in 2011, nor was there an association between the 2La inversion and infection with Plasmodium

Temperature during larval development and adult maintenance influences the survival of Anopheles gambiae s.s.

PubMed

Christiansen-Jucht, Céline; Parham, Paul E; Saddler, Adam; Koella, Jacob C; Basáñez, María-Gloria

2014-11-05

Malaria transmission depends on vector life-history parameters and population dynamics, and particularly on the survival of adult Anopheles mosquitoes. These dynamics are sensitive to climatic and environmental factors, and temperature is a particularly important driver. Data currently exist on the influence of constant and fluctuating adult environmental temperature on adult Anopheles gambiae s.s. survival and on the effect of larval environmental temperature on larval survival, but none on how larval temperature affects adult life-history parameters. Mosquito larvae and pupae were reared individually at different temperatures (23 ± 1°C, 27 ± 1°C, 31 ± 1°C, and 35 ± 1°C), 75 ± 5% relative humidity. Upon emergence into imagoes, individual adult females were either left at their larval temperature or placed at a different temperature within the range above. Survival was monitored every 24 hours and data were analysed using non-parametric and parametric methods. The Gompertz distribution fitted the survivorship data better than the gamma, Weibull, and exponential distributions overall and was adopted to describe mosquito mortality rates. Increasing environmental temperature during the larval stages decreased larval survival (p < 0.001). Increases of 4°C (from 23°C to 27°C, 27°C to 31°C, and 31°C to 35°C), 8°C (27°C to 35°C) and 12°C (23°C to 35°C) statistically significantly increased larval mortality (p < 0.001). Higher environmental temperature during the adult stages significantly lowered adult survival overall (p < 0.001), with increases of 4°C and 8°C significantly influencing survival (p < 0.001). Increasing the larval environment temperature also significantly increased adult mortality overall (p < 0.001): a 4°C increase (23°C to 27°C) did not significantly affect adult survival (p > 0.05), but an 8°C increase did (p < 0.05). The effect of a 4°C increase in larval temperature from 27

Pyrethroid resistance in Anopheles gambiae leads to increased susceptibility to the entomopathogenic fungi Metarhizium anisopliae and Beauveria bassiana

PubMed Central

2010-01-01

Background Entomopathogenic fungi are being investigated as a new mosquito control tool because insecticide resistance is preventing successful mosquito control in many countries, and new methods are required that can target insecticide-resistant malaria vectors. Although laboratory studies have previously examined the effects of entomopathogenic fungi against adult mosquitoes, most application methods used cannot be readily deployed in the field. Because the fungi are biological organisms it is important to test potential field application methods that will not adversely affect them. The two objectives of this study were to investigate any differences in fungal susceptibility between an insecticide-resistant and insecticide-susceptible strain of Anopheles gambiae sensu stricto, and to test a potential field application method with respect to the viability and virulence of two fungal species Methods Pieces of white polyester netting were dipped in Metarhizium anisopliae ICIPE-30 or Beauveria bassiana IMI391510 mineral oil suspensions. These were kept at 27 ± 1°C, 80 ± 10% RH and the viability of the fungal conidia was recorded at different time points. Tube bioassays were used to infect insecticide-resistant (VKPER) and insecticide-susceptible (SKK) strains of An. gambiae s.s., and survival analysis was used to determine effects of mosquito strain, fungus species or time since fungal treatment of the net. Results The resistant VKPER strain was significantly more susceptible to fungal infection than the insecticide-susceptible SKK strain. Furthermore, B. bassiana was significantly more virulent than M. anisopliae for both mosquito strains, although this may be linked to the different viabilities of these fungal species. The viability of both fungal species decreased significantly one day after application onto polyester netting when compared to the viability of conidia remaining in suspension. Conclusions The insecticide-resistant mosquito strain was susceptible

Gene expression divergence between malaria vector sibling species Anopheles gambiae and An. coluzzii from rural and urban Yaoundé Cameroon

PubMed Central

Cassone, Bryan J.; Kamdem, Colince; Cheng, Changde; Tan, John C.; Hahn, Matthew W.; Costantini, Carlo; Besansky, Nora J.

2014-01-01

Divergent selection based on aquatic larval ecology is a likely factor in the recent isolation of two broadly sympatric and morphologically identical African mosquito species, the malaria vectors Anopheles gambiae and An. coluzzii. Population-based genome scans have revealed numerous candidate regions of recent positive selection, but have provided few clues as to the genetic mechanisms underlying behavioral and physiological divergence between the two species, phenotypes which themselves remain obscure. To uncover possible genetic mechanisms, we compared global transcriptional profiles of natural and experimental populations using gene-based microarrays. Larvae were sampled as second and fourth instars from natural populations in and around the city of Yaoundé, capital of Cameroon, where the two species segregate along a gradient of urbanization. Functional enrichment analysis of differentially expressed genes revealed that An. coluzzii—the species that breeds in more stable, biotically complex and potentially polluted urban water bodies—over-expresses genes implicated in detoxification and immunity relative to An. gambiae, which breeds in more ephemeral and relatively depauperate pools and puddles in suburbs and rural areas. Moreover, our data suggest that such over-expression by An. coluzzii is not a transient result of induction by xenobiotics in the larval habitat, but an inherent and presumably adaptive response to repeatedly encountered environmental stressors. Finally, we find no significant overlap between the differentially expressed loci and previously identified genomic regions of recent positive selection, suggesting that transcriptome divergence is regulated by trans-acting factors rather than cis-acting elements. PMID:24673723

Reduced susceptibility to DDT in field populations of Anopheles quadriannulatus and Anopheles arabiensis in Malawi: evidence for larval selection.

PubMed

Mzilahowa, T; Ball, A J; Bass, C; Morgan, J C; Nyoni, B; Steen, K; Donnelly, M J; Wilding, C S

2008-09-01

Bioassays for insecticide resistance in adult mosquitoes were conducted on samples of Anopheles gambiae Giles s.l. (Diptera: Culicidae) species collected as larvae from breeding sites in the lower Shire Valley, Malawi. The results indicate full susceptibility to permethrin, deltamethrin and malathion, but reduced susceptibility to DDT in one sample from Thom (LT(50) of 8.39 min for females and 25.09 min for males). Polymerase chain reaction-based species identification of the mosquitoes assayed revealed a mixture of Anopheles arabiensis Patton and Anopheles quadriannulatus (Theobold). The LT(50) did not differ significantly between species. Genotyping of the L1014F and L1014S kdr alleles showed all mosquito specimens to be homozygous wild type; thus the reduced susceptibility detected is not attributable to target site insensitivity and instead is likely to be metabolic in nature. Anopheles quadriannulatus is characteristically zoophagic and exophilic. Indeed, of 82 Anopheles collected through knockdown collections within dwellings, only one was An. quadriannulatus and the rest were An. arabiensis. They are unlikely, therefore, to have been exposed to selection pressure arising from insecticide-treated net usage or to DDT indoor residual spraying. Therefore, it is suggested that this example of reduced susceptibility to DDT in An. quadriannulatus reflects selection in the larval stages.

Activity of Cinnamomum osmophloeum leaf essential oil against Anopheles gambiae s.s

PubMed Central

2014-01-01

Background The increasing status of insecticide resistant mosquitoes in sub-Saharan Africa is a threatening alert to the existing control efforts. All sibling species of An. gambiae complex have evolved insecticide resistance in wild populations for different approved classes of the insecticides currently in use in the field. An alternative compound for vector control is absolutely urgently needed. In this study, the larvicidal activity and chemical composition of the Cinnamomum osmophloeum leaf essential oils were investigated. Methods C. osmophloeum leaf essential oils were extracted by hydrodistillation in a Clevenger-type apparatus for 6 hours, and their chemical compositions identified using GC-MS. These oils were evaluated against An. gambiae s.s. in both laboratory and semi-field situations. The WHO test procedures for monitoring larvicidal efficacy in malaria vectors were used. Results The composition of C. osmophloeum leaf essential oil has been found to have 11 active compounds. The most abundant compound was trans-cinnamaldehyde (70.20%) and the least abundant was caryophyllene oxide (0.08%). The larvicidal activity was found to be dosage and time dependant both in laboratory and semi-field environments with mortality ranging from 0% to 100%. The LC50 value was found to vary from 22.18 to 58.15 μg/ml in the laboratory while in semi-field environments it was 11.91 to 63.63 μg/ml. The LC90 value was found to range between 57.71 to 91.54 μg/ml in the laboratory while in semi-field environments was 52.07 to 173.77 μg/ml. Mortality ranged from 13% to 100% in the laboratory while in semi-field environments it ranged between 43% to 100% within mortality recording time intervals of 12, 24, 48, and 72 hours. Conclusions The larvicidal activity shown by C. osmophloeum leaf essential oil is a promising alternative to existing larvicides or to be incorporated in integrated larval source management compounds for An. gambiae s.s control. The efficacy

[Preliminary evaluation of the insecticide susceptibility in Anopheles gambiae and Culex quinquefasciatus from Lobito (Angola), using WHO standard assay].

PubMed

Toto, J C; Besnard, P; Le Mire, J; Almeida, D S I; Dos Santos, M A; Fortes, F; Foumane, V; Simard, F; Awono-Ambene, H P; Carnevale, P

2011-10-01

Field collections of the most common urban mosquito vectors Anopheles gambiae and Culex quinquefasciatus were carried out in June 2003, March 2004 and November 2005 to gather preliminary data on the insecticide susceptibility in mosquitoes from Lobito (Angola) using the WHO standard bioassays. Bioassays were performed on F0 adults emerging from the field larval collections and on unfed adults from landing catches on volunteers. Batches of mosquitoes from three selected locations (Alto Liro, San Jao and Bela Vista) were exposed for 1 hour to several insecticides such as DDT 4%, carbosulfan 0.4%, permethrin 1%, deltamethrin 0.05% and cyfluthrin 0.15%, in order to estimate the immediate knockdown times (kdT50 and kdT95) and the mortality rate after exposure. The results revealed that mosquito susceptibility to insecticides varied depending on the insecticide, the site and the period of collection. The main local malaria vector A. gambiae (both M and S forms) was basically resistant to DDT and susceptible to all pyrethoids, regardless of the period and the site of collections. The overall mortality rate due to DDT was 73% in Alto Liro, 89% in San Jao and varied depending on the period in Bela Vista between 95% in March 2004 and 100% in November 2005. The mortality due to pyrethoids was 100% at all locations, with the kdT50 and KdT95 times ranging between 9 and 16 minutes and between 18 and 29 minutes, respectively. Concerning the C. quinquefasciatus, populations from Yard and Caponte were resistant to all insecticides tested; the mortality rate was 40% with deltamethrin and 70% with permethrin, while no lethal effect was observed with DDT or carbosulfan. In conclusion, despite its probable high resistance to DDT, the main local malaria vector A. gambiae remained fully susceptible to pyrethroids. This could forecast a good biological efficacy of the scheduled vector control interventions in Angola, based on a large-scale distribution of long-lasting, insecticide

Genetic sex separation in Anopheles arabiensis and the production of sterile hybrids.

PubMed

Curtis, C F

1978-01-01

The gene for dieldrin resistance has been artificially male-linked so that females can be selectively killed with dieldrin. By intercrossing different sibling species of the Anopheles gambiae complex, batches consisting of sterile males only can be reared. This seems to have potential for use in genetic control operations.

Age and prior blood feeding of Anopheles gambiae influences their susceptibility and gene expression patterns to ivermectin-containing blood meals.

PubMed

Seaman, Jonathan A; Alout, Haoues; Meyers, Jacob I; Stenglein, Mark D; Dabiré, Roch K; Lozano-Fuentes, Saul; Burton, Timothy A; Kuklinski, Wojtek S; Black, William C; Foy, Brian D

2015-10-15

Ivermectin has been proposed as a novel malaria transmission control tool based on its insecticidal properties and unique route of acquisition through human blood. To maximize ivermectin's effect and identify potential resistance/tolerance mechanisms, it is important to understand its effect on mosquito physiology and potential to shift mosquito population age-structure. We therefore investigated ivermectin susceptibility and gene expression changes in several age groups of female Anopheles gambiae mosquitoes. The effect of aging on ivermectin susceptibility was analyzed in three age groups (2, 6, and 14-days) of colonized female Anopheles gambiaemosquitoes using standard survivorship assays. Gene expression patterns were then analyzed by transcriptome sequencing on an Illumina HiSeq 2500 platform. RT-qPCR was used to validate transcriptional changes and also to examine expression in a different, colonized strain and in wild mosquitoes, both of which blood fed naturally on an ivermectin-treated person. Mosquitoes of different ages and blood meal history died at different frequencies after ingesting ivermectin. Mortality was lowest in 2-day old mosquitoes exposed on their first blood meal and highest in 6-day old mosquitoes exposed on their second blood meal. Twenty-four hours following ivermectin ingestion, 101 and 187 genes were differentially-expressed relative to control blood-fed, in 2 and 6-day groups, respectively. Transcription patterns of select genes were similar in membrane-fed, colonized, and naturally-fed wild vectors. Transcripts from several unexpected functional classes were highly up-regulated, including Niemann-Pick Type C (NPC) genes, peritrophic matrix-associated genes, and immune-response genes, and these exhibited different transcription patterns between age groups, which may explain the observed susceptibility differences. Niemann-Pick Type 2 genes were the most highly up-regulated transcripts after ivermectin ingestion (up to 160 fold) and

Evaluation of the long-term efficacy of K-Othrine® PolyZone on three surfaces against laboratory reared Anopheles gambiae in semi-field conditions.

PubMed

Dunford, James C; Estep, Alden S; Waits, Christy M; Richardson, Alec G; Hoel, David F; Horn, Karin; Walker, Todd W; Blersch, Jessika S; Kerce, Jerry D; Wirtz, Robert A

2018-02-23

In this semi-field study, a new polymer-enhanced deltamethrin formulation, K-Othrine ® PolyZone, was compared to a standard deltamethrin product for residual activity against a susceptible strain of laboratory-reared Anopheles gambiae using standard WHO cone bioassays. Residual insecticide efficacy was recorded after exposure to metal, cement and wood panels maintained in experimental huts in sub-tropical environmental conditions in north central Florida, USA, and panels stored in a climate controlled chamber located at the Centers for Disease Control and Prevention, Georgia, USA. K-Othrine ® PolyZone demonstrated 100% control on metal and cement panels 1 year post application and > 80% control on wood panels up to 6 mo. The new formulation should be considered for use in indoor residual spray programmes requiring long-term control of malaria vectors.

Parallel evolution or purifying selection, not introgression, explain similarity in the pyrethroid detoxification linked GSTE4 of Anopheles gambiae and An. arabiensis

PubMed Central

Wilding, C.S.; Weetman, D.; Rippon, E.J.; Steen, K.; Mawejje, H.D.; Barsukov, I.; Donnelly, M.J.

2014-01-01

Insecticide resistance is a major impediment to the control of vectors and pests of public health importance and is a strongly selected trait capable of rapid spread, sometimes even between closely-related species. Elucidating the mechanisms generating insecticide resistance in mosquito vectors of disease, and understanding the spread of resistance within and between populations and species are vital for the development of robust resistance management strategies. Here we studied the mechanisms of resistance in two sympatric members of the Anopheles gambiae species complex – the major vector of malaria in sub-Saharan Africa – in order to understand how resistance has developed and spread in eastern Uganda, a region with some of the highest levels of malaria. In eastern Uganda, where the mosquitoes Anopheles arabiensis and An. gambiae can be found sympatrically, low levels of hybrids (0.4%) occur, offering a route for introgression of adaptively important variants between species. In independent microarray studies of insecticide resistance, Gste4, an insect-specific glutathione S-transferase, was among the most significantly up-regulated genes in both species. To test the hypothesis of interspecific introgression, we sequenced 2.3kbp encompassing Gste4. Whilst this detailed sequencing ruled out introgression, we detected strong positive selection acting on Gste4. However, these sequences, followed by haplotype-specific qPCR, showed that the apparent up-regulation in An. arabiensis is a result of allelic variation across the microarray probe binding sites which artefactually elevates the gene expression signal. Thus, face-value acceptance of microarray data can be misleading and it is advisable to conduct a more detailed investigation of the causes and nature of such signal. The identification of positive selection acting on this locus led us to functionally express and characterise allelic variants of GSTE4. Although the in vitro data do not support a direct role

Genetic sex separation in Anopheles arabiensis and the production of sterile hybrids

PubMed Central

Curtis, C. F.

1978-01-01

The gene for dieldrin resistance has been artificially male-linked so that females can be selectively killed with dieldrin. By intercrossing different sibling species of the Anopheles gambiae complex, batches consisting of sterile males only can be reared. This seems to have potential for use in genetic control operations. PMID:308410

Chlorfenapyr (A Pyrrole Insecticide) Applied Alone or as a Mixture with Alpha-Cypermethrin for Indoor Residual Spraying against Pyrethroid Resistant Anopheles gambiae sl: An Experimental Hut Study in Cove, Benin

PubMed Central

Ngufor, Corine; Critchley, Jessica; Fagbohoun, Josias; N’Guessan, Raphael; Todjinou, Damien; Rowland, Mark

2016-01-01

Background Indoor spraying of walls and ceilings with residual insecticide remains a primary method of malaria control. Insecticide resistance in malaria vectors is a growing problem. Novel insecticides for indoor residual spraying (IRS) which can improve the control of pyrethroid resistant malaria vectors are urgently needed. Insecticide mixtures have the potential to improve efficacy or even to manage resistance in some situations but this possibility remains underexplored experimentally. Chlorfenapyr is a novel pyrrole insecticide which has shown potential to improve the control of mosquitoes which are resistant to current WHO-approved insecticides. Method The efficacy of IRS with chlorfenapyr applied alone or as a mixture with alpha-cypermeththrin (a pyrethroid) was evaluated in experimental huts in Cove, Southern Benin against wild free flying pyrethroid resistant Anopheles gambiae sl. Comparison was made with IRS with alpha-cypermethrin alone. Fortnightly 30-minute in situ cone bioassays were performed to assess the residual efficacy of the insecticides on the treated hut walls. Results Survival rates of wild An gambiae from the Cove hut site in WHO resistance bioassays performed during the trial were >90% with permethrin and deltamethrin treated papers. Mortality of free-flying mosquitoes entering the experimental huts was 4% in the control hut. Mortality with alpha-cypermethrin IRS did not differ from the control (5%, P>0.656). The highest mortality was achieved with chlorfenapyr alone (63%). The alpha-cypermethrin + chlorfenapyr mixture killed fewer mosquitoes than chlorfenapyr alone (43% vs. 63%, P<0.001). While the cone bioassays showed a more rapid decline in residual mortality with chlorfenapyr IRS to <30% after only 2 weeks, fortnightly mortality rates of wild free-flying An gambiae entering the chlorfenapyr IRS huts were consistently high (50–70%) and prolonged, lasting over 4 months. Conclusion IRS with chlorfenapyr shows potential to

Dissecting the organ specificity of insecticide resistance candidate genes in Anopheles gambiae: known and novel candidate genes.

PubMed

Ingham, Victoria A; Jones, Christopher M; Pignatelli, Patricia; Balabanidou, Vasileia; Vontas, John; Wagstaff, Simon C; Moore, Jonathan D; Ranson, Hilary

2014-11-25

The elevated expression of enzymes with insecticide metabolism activity can lead to high levels of insecticide resistance in the malaria vector, Anopheles gambiae. In this study, adult female mosquitoes from an insecticide susceptible and resistant strain were dissected into four different body parts. RNA from each of these samples was used in microarray analysis to determine the enrichment patterns of the key detoxification gene families within the mosquito and to identify additional candidate insecticide resistance genes that may have been overlooked in previous experiments on whole organisms. A general enrichment in the transcription of genes from the four major detoxification gene families (carboxylesterases, glutathione transferases, UDP glucornyltransferases and cytochrome P450s) was observed in the midgut and malpighian tubules. Yet the subset of P450 genes that have previously been implicated in insecticide resistance in An gambiae, show a surprisingly varied profile of tissue enrichment, confirmed by qPCR and, for three candidates, by immunostaining. A stringent selection process was used to define a list of 105 genes that are significantly (p ≤0.001) over expressed in body parts from the resistant versus susceptible strain. Over half of these, including all the cytochrome P450s on this list, were identified in previous whole organism comparisons between the strains, but several new candidates were detected, notably from comparisons of the transcriptomes from dissected abdomen integuments. The use of RNA extracted from the whole organism to identify candidate insecticide resistance genes has a risk of missing candidates if key genes responsible for the phenotype have restricted expression within the body and/or are over expression only in certain tissues. However, as transcription of genes implicated in metabolic resistance to insecticides is not enriched in any one single organ, comparison of the transcriptome of individual dissected body parts cannot

Modeling the Response of Anopheles gambiae (Diptera: Culicidae) Populations in the Kenya Highlands to a Rise in Mean Annual Temperature.

PubMed

Wallace, Dorothy; Prosper, Olivia; Savos, Jacob; Dunham, Ann M; Chipman, Jonathan W; Shi, Xun; Ndenga, Bryson; Githeko, Andrew

2017-03-01

A dynamical model of Anopheles gambiae larval and adult populations is constructed that matches temperature-dependent maturation times and mortality measured experimentally as well as larval instar and adult mosquito emergence data from field studies in the Kenya Highlands. Spectral classification of high-resolution satellite imagery is used to estimate household density. Indoor resting densities collected over a period of one year combined with predictions of the dynamical model give estimates of both aquatic habitat and total adult mosquito densities. Temperature and precipitation patterns are derived from monthly records. Precipitation patterns are compared with average and extreme habitat estimates to estimate available aquatic habitat in an annual cycle. These estimates are coupled with the original model to produce estimates of adult and larval populations dependent on changing aquatic carrying capacity for larvae and changing maturation and mortality dependent on temperature. This paper offers a general method for estimating the total area of aquatic habitat in a given region, based on larval counts, emergence rates, indoor resting density data, and number of households.Altering the average daily temperature and the average daily rainfall simulates the effect of climate change on annual cycles of prevalence of An. gambiae adults. We show that small increases in average annual temperature have a large impact on adult mosquito density, whether measured at model equilibrium values for a single square meter of habitat or tracked over the course of a year of varying habitat availability and temperature. © The Authors 2016. Published by Oxford University Press on behalf of Entomological Society of America. All rights reserved. For Permissions, please email: journals.permissions@oup.com.

Biophysical Analysis of Anopheles gambiae Leucine-Rich Repeat Proteins APL1A1, APL1B and APL1C and Their Interaction with LRIM1

DOE PAGES

Williams, Marni; Summers, Brady J.; Baxter, Richard H. G.; ...

2015-03-16

Natural infection of Anopheles gambiae by malaria-causing Plasmodium parasites is significantly influenced by the APL1 genetic locus. The locus contains three closely related leucine-rich repeat (LRR) genes, APL1A, APL1B and APL1C. Multiple studies have reported the participation of APL1A—C in the immune response of A. gambiae to invasion by both rodent and human Plasmodium isolates. APL1C forms a heterodimer with the related LRR protein LRIM1 via a C-terminal coiled-coil domain that is also present in APL1A and APL1B. The LRIM1/APL1C heterodimer protects A. gambiae from infection by binding the complement-like protein TEP1 to form a stable and active immune complex.more » We report solution x-ray scatting data for the LRIM1/APL1C heterodimer, the oligomeric state of LRIM1/APL1 LRR domains in solution and the crystal structure of the APL1B LRR domain. The LRIM1/APL1C heterodimeric complex has a flexible and extended structure in solution. In contrast to the APL1A, APL1C and LRIM1 LRR domains, the APL1B LRR domain is a homodimer. The crystal structure of APL1B-LRR shows that the homodimer is formed by an N-terminal helix that complements for the absence of an N-terminal capping motif in APL1B, which is a unique distinction within the LRIM1/APL1 protein family. Full-length APL1A 1 and APL1B form a stable complex with LRIM1. Our results support a model in which APL1A 1, APL1B and APL1C can all form an extended, flexible heterodimer with LRIM1, providing a repertoire of functional innate immune complexes to protect A. gambiae from a diverse array of pathogens.« less

Biophysical Analysis of Anopheles gambiae Leucine-Rich Repeat Proteins APL1A1, APL1B and APL1C and Their Interaction with LRIM1

DOE Office of Scientific and Technical Information (OSTI.GOV)

Williams, Marni; Summers, Brady J.; Baxter, Richard H. G.

Natural infection of Anopheles gambiae by malaria-causing Plasmodium parasites is significantly influenced by the APL1 genetic locus. The locus contains three closely related leucine-rich repeat (LRR) genes, APL1A, APL1B and APL1C. Multiple studies have reported the participation of APL1A—C in the immune response of A. gambiae to invasion by both rodent and human Plasmodium isolates. APL1C forms a heterodimer with the related LRR protein LRIM1 via a C-terminal coiled-coil domain that is also present in APL1A and APL1B. The LRIM1/APL1C heterodimer protects A. gambiae from infection by binding the complement-like protein TEP1 to form a stable and active immune complex.more » We report solution x-ray scatting data for the LRIM1/APL1C heterodimer, the oligomeric state of LRIM1/APL1 LRR domains in solution and the crystal structure of the APL1B LRR domain. The LRIM1/APL1C heterodimeric complex has a flexible and extended structure in solution. In contrast to the APL1A, APL1C and LRIM1 LRR domains, the APL1B LRR domain is a homodimer. The crystal structure of APL1B-LRR shows that the homodimer is formed by an N-terminal helix that complements for the absence of an N-terminal capping motif in APL1B, which is a unique distinction within the LRIM1/APL1 protein family. Full-length APL1A 1 and APL1B form a stable complex with LRIM1. Our results support a model in which APL1A 1, APL1B and APL1C can all form an extended, flexible heterodimer with LRIM1, providing a repertoire of functional innate immune complexes to protect A. gambiae from a diverse array of pathogens.« less

Field experiments of Anopheles gambiae attraction to local fruits/seedpods and flowering plants in Mali to optimize strategies for malaria vector control in Africa using attractive toxic sugar bait methods

PubMed Central

2010-01-01

Background Based on recent studies in Israel demonstrating that attractive toxic sugar bait (ATSB) methods can be used to decimate local anopheline and culicine mosquito populations, an important consideration is whether the same methods can be adapted and improved to attract and kill malaria vectors in Africa. The ATSB approach uses fruit or flower scent as an attractant, sugar solution as a feeding stimulant, and an oral toxin. The ATSB solutions are either sprayed on vegetation or suspended in simple bait stations, and the mosquitoes ingesting the toxic solutions are killed. As such, this approach targets sugar-feeding female and male mosquitoes. This study examines the attractiveness of African malaria vectors to local fruits/seedpods and flowering plants, key biological elements of the ATSB approach for mosquito control. Methods Three field experiments were conducted at sites in Mali. The attraction of Anopheles gambiae s.l. to 26 different local fruits and seedpods was determined at a site in the semi-arid Bandiagara District of Mali. Wire mesh glue traps with fruits/seedpods suspended on skewers inside were set along a seasonal lagoon. Seven replicates of each fruit/seedpod species were tested, with a water-soaked sponge and a sugar-soaked sponge as controls. The attraction of An. gambiae s.l. to 26 different types of flowering plants was determined at a site near Mopti in Mali. The flowering plants held in a water-filled buried container were tested using the same glue traps, with controls including water only and sugar solution. Six replicates of each selected plant type were tested on transects between rice paddies. Additional studies using CDC light traps were done to determine the relative densities and periodicity of An. gambiae s.l. attraction to branches of the most highly attractive flowering plant, branches without flowers, human odor, and candescent light. Results Of the 26 fruits and seedpods tested, 6 were attractive to An. gambiae s.l. females

Salivary Polytene Chromosome Map of Anopheles darlingi, the Main Vector of Neotropical Malaria

PubMed Central

Rafael, Míriam S.; Rohde, Cláudia; Bridi, Letícia C.; da Silva Valente Gaiesky, Vera Lúcia; Tadei, Wanderli P.

2010-01-01

New photomap of Anopheles (Nyssorhynchus) darlingi Root, 1926, is described for a population from Guajará-Mirim, State of Rondonia, Brazil. The number of sections in the previous A. darlingi reference map was maintained and new subsections were added to the five chromosome arms. Breakage points of paracentric inversions had been previously incorporated into the photomap of this species. An additional inversion is reported, called 3Lc, totaling 14 inversions in the A. darlingi chromosome arms. The proposed photomap is potentially useful for further evolutionary studies in addition to physical and in silico chromosome mapping using A. darlingi genomic and transcriptome sequences. Furthermore, in our attempt to compare sections of the 2R chromosome arm of A. darlingi with Anopheles funestus, Anopheles stephensi, and Anopheles gambiae, we found great differences in the arrangement of the polytene chromosome bands, which are consistent with the known phylogenetic divergence of these species. PMID:20682862

The dominant Anopheles vectors of human malaria in Africa, Europe and the Middle East: occurrence data, distribution maps and bionomic précis

PubMed Central

2010-01-01

Background This is the second in a series of three articles documenting the geographical distribution of 41 dominant vector species (DVS) of human malaria. The first paper addressed the DVS of the Americas and the third will consider those of the Asian Pacific Region. Here, the DVS of Africa, Europe and the Middle East are discussed. The continent of Africa experiences the bulk of the global malaria burden due in part to the presence of the An. gambiae complex. Anopheles gambiae is one of four DVS within the An. gambiae complex, the others being An. arabiensis and the coastal An. merus and An. melas. There are a further three, highly anthropophilic DVS in Africa, An. funestus, An. moucheti and An. nili. Conversely, across Europe and the Middle East, malaria transmission is low and frequently absent, despite the presence of six DVS. To help control malaria in Africa and the Middle East, or to identify the risk of its re-emergence in Europe, the contemporary distribution and bionomics of the relevant DVS are needed. Results A contemporary database of occurrence data, compiled from the formal literature and other relevant resources, resulted in the collation of information for seven DVS from 44 countries in Africa containing 4234 geo-referenced, independent sites. In Europe and the Middle East, six DVS were identified from 2784 geo-referenced sites across 49 countries. These occurrence data were combined with expert opinion ranges and a suite of environmental and climatic variables of relevance to anopheline ecology to produce predictive distribution maps using the Boosted Regression Tree (BRT) method. Conclusions The predicted geographic extent for the following DVS (or species/suspected species complex*) is provided for Africa: Anopheles (Cellia) arabiensis, An. (Cel.) funestus*, An. (Cel.) gambiae, An. (Cel.) melas, An. (Cel.) merus, An. (Cel.) moucheti and An. (Cel.) nili*, and in the European and Middle Eastern Region: An. (Anopheles) atroparvus, An. (Ano

Experimental hut evaluation of bednets treated with an organophosphate (chlorpyrifos-methyl) or a pyrethroid (lambdacyhalothrin) alone and in combination against insecticide-resistant Anopheles gambiae and Culex quinquefasciatus mosquitoes

PubMed Central

Asidi, Alex N; N' Guessan, Raphael; Koffi, Alphonsine A; Curtis, Christopher F; Hougard, Jean-Marc; Chandre, Fabrice; Corbel, Vincent; Darriet, Frédéric; Zaim, Morteza; Rowland, Mark W

2005-01-01

Background Pyrethroid resistant mosquitoes are becoming increasingly common in parts of Africa. It is important to identify alternative insecticides which, if necessary, could be used to replace or supplement the pyrethroids for use on treated nets. Certain compounds of an earlier generation of insecticides, the organophosphates may have potential as net treatments. Methods Comparative studies of chlorpyrifos-methyl (CM), an organophosphate with low mammalian toxicity, and lambdacyhalothrin (L), a pyrethroid, were conducted in experimental huts in Côte d'Ivoire, West Africa. Anopheles gambiae and Culex quinquefasciatus mosquitoes from the area are resistant to pyrethroids and organophosphates (kdr and insensitive acetylcholinesterase Ace.1R). Several treatments and application rates on intact or holed nets were evaluated, including single treatments, mixtures, and differential wall/ceiling treatments. Results and Conclusion All of the treatments were effective in reducing blood feeding from sleepers under the nets and in killing both species of mosquito, despite the presence of the kdr and Ace.1R genes at high frequency. In most cases, the effects of the various treatments did not differ significantly. Five washes of the nets in soap solution did not reduce the impact of the insecticides on A. gambiae mortality, but did lead to an increase in blood feeding. The three combinations performed no differently from the single insecticide treatments, but the low dose mixture performed encouragingly well indicating that such combinations might be used for controlling insecticide resistant mosquitoes. Mortality of mosquitoes that carried both Ace.1R and Ace.1S genes did not differ significantly from mosquitoes that carried only Ace.1S genes on any of the treated nets, indicating that the Ace.1R allele does not confer effective resistance to chlorpyrifos-methyl under the realistic conditions of an experimental hut. PMID:15918909

The interplay between dose and immune system activation determines fungal infection outcome in the African malaria mosquito, Anopheles gambiae.

PubMed

Rhodes, Victoria L; Thomas, Matthew B; Michel, Kristin

2018-08-01

The Toll pathway is a central regulator of antifungal immunity in insects. In mosquitoes, the Toll pathway affects infections with the fungal entomopathogen, Beauveria bassiana, which is considered a potential mosquito biopesticide. We report here the use of B. bassiana strain I93-825 in Anopheles gambiae to analyze the impact of Toll pathway modulation on mosquito survival. Exposure to a narrow dose range of conidia by direct contact decreased mosquito longevity and median survival. In addition, fungal exposure dose correlated positively and linearly with hazard ratio. Increased Toll signaling by knockdown of its inhibitor, cactus, decreased survivorship of uninfected females, increased mosquito survival after low dose B. bassiana exposure, but had little effect following exposure to higher doses. This observed trade-off could have implications for development of B. bassiana as a prospective vector control tool. On the one hand, selection for small increases in mosquito immune signaling across a narrow dose range could impair efficacy of B. bassiana. On the other hand, costs of immunity and the capacity for higher doses of fungus to overwhelm immune responses could limit evolution of resistance. Copyright © 2018 Elsevier Ltd. All rights reserved.

Vectorial status and insecticide resistance of Anopheles funestus from a sugar estate in southern Mozambique.

PubMed

Kloke, R Graham; Nhamahanga, Eduardo; Hunt, Richard H; Coetzee, Maureen

2011-02-09

The dual problems of rising insecticide resistance in the malaria vectors and increasing human malaria cases since 2001 in southern Mozambique are cause for serious concern. The selection of insecticides for use in indoor residual spraying (IRS) programmes is highly dependent on the extent to which local mosquitoes are susceptible to the approved classes of insecticides. The insecticide resistance status and role in malaria transmission of Anopheles funestus was evaluated at the Maragra Sugar Estate in southern Mozambique where an IRS vector control programme has been in operation for seven years using the carbamate insecticide bendiocarb. No Anopheles species were captured inside the sugar estate control area. Anopheles funestus group captured outside of the estate represented 90% (n = 475) of the total collections. Of the specimens identified to species by PCR (n = 167), 95% were An. funestus s.s. One An. rivulorum was identified and seven specimens did not amplify. The Anopheles gambiae complex was less abundant (n = 53) and of those identified (n = 33) 76% were An. arabiensis and 24% An. merus. Insecticide susceptibility tests showed that wild-caught and F-1 family An. funestus were resistant to deltamethrin (32.5% mortality) and lambda-cyhalothrin (14.6% mortality), less so to bendiocarb (71.5% mortality) and fully susceptible to both malathion and DDT (100%). Bendiocarb and pyrethroid resistance was nullified using 4% piperonyl butoxide (Pbo), strongly suggesting that both are mediated by P450 monooxygenase detoxification. ELISA tests of An. funestus for Plasmodium falciparum, gave a sporozoite rate of 6.02% (n = 166). One unidentified member of the An. gambiae complex tested positive for P. falciparum sporozoites. Anopheles funestus was found to be the most abundant and principle vector of malaria in this area, with members of the An. gambiae complex being secondary vectors. Despite the continual use of bendiocarb within the estate for seven years and the

Efficacy of PermaNet® 3.0 and PermaNet® 2.0 nets against laboratory-reared and wild Anopheles gambiae sensu lato populations in northern Tanzania.

PubMed

Kweka, Eliningaya J; Lyaruu, Lucile J; Mahande, Aneth M

2017-01-18

Mosquitoes have developed resistance against pyrethroids, the only class of insecticides approved for use on long-lasting insecticidal nets (LLINs). The present study sought to evaluate the efficacy of the pyrethroid synergist PermaNet® 3.0 LLIN versus the pyrethroid-only PermaNet® 2.0 LLIN, in an East African hut design in Lower Moshi, northern Tanzania. In this setting, resistance to pyrethroid insecticides has been identified in Anopheles gambiae mosquitoes. Standard World Health Organization bioefficacy evaluations were conducted in both laboratory and experimental huts. Experimental hut evaluations were conducted in an area where there was presence of a population of highly pyrethroid-resistant An. arabiensis mosquitoes. All nets used were subjected to cone bioassays and then to experimental hut trials. Mosquito mortality, blood-feeding inhibition and personal protection rate were compared between untreated nets, unwashed LLINs and LLINs that were washed 20 times. Both washed and unwashed PermaNet® 2.0 and PermaNet® 3.0 LLINs had knockdown and mortality rates of 100% against a susceptible strain of An. gambiae sensu stricto. The adjusted mortality rate of the wild mosquito population after use of the unwashed PermaNet® 3.0 and PermaNet® 2.0 nets was found to be higher than after use of the washed PermaNet® 2.0 and PermaNet® 3.0 nets. Given the increasing incidence of pyrethroid resistance in An. gambiae mosquitoes in Tanzania, we recommend that consideration is given to its distribution in areas with pyrethroid-resistant malaria vectors within the framework of a national insecticide-resistance management plan.

Larvicidal effect of aqueous and ethanolic extracts of Senna alata on Anopheles gambiae, Culex quinquefasciatus and Aedes aegypti.

PubMed

Edwin, Ubulom Peace Mayen; Nyiutaha, Imandeh Godwin; Essien, Akpabio Eno; Nnamdi, Opara Kenneth; Sunday, Ekanem Mfon

2013-05-01

Senna alata is locally used in South Eastern Nigeria in the treatment of several infections which include ringworm and other parasitic skin diseases.The larvicidal activities of aqueous and ethanolic leaf and stem extracts of S. alata were evaluated in static bioassays, on fourth instar larvae of Anopheles gambiae, Culex quinquefasciatus and Aedes aegypti, at extract concentrations of 0.15, 0.30, 0.45, 0.60 and 0.75% w/v, for 72 hours. Mortality of larvae exposed to the different extracts increased with increase in extract concentration and time of exposure. This study revealed a differential potency of the extracts used and a difference in susceptibility of larvae to the extracts as evident by the 72hLC₅₀ values obtained. The leaf extract proved to be more lethal to the larvae than the stem extract as judged by the 72hLC₅₀ values obtained both for the aqueous as well as the ethanolic extracts assayed. Phytochemical screening of the plant parts investigated revealed the presence of some plant metabolites, which have been reported in separate studies to be lethal to mosquito larvae. Results obtained from this study suggest that the leaf and stem extracts of S. alata possess a promising larvicidal potential which can be exploited in mosquito vector control.

Eye pigments in wild-type and eye-color mutant strains of the African malaria vector Anopheles gambiae.

PubMed

Beard, C B; Benedict, M Q; Primus, J P; Finnerty, V; Collins, F H

1995-01-01

Chromatographic analysis of pigments extracted from wild-type eyes of the mosquito Anopheles gambiae reveals the presence of the ommatin precursor 3-hydroxykynurenine, its transamination derivative xanthurenic acid, and a dark, red-brown pigment spot that probably is composed of two or more low mobility xanthommatins. No colored or fluorescent pteridines are evident. Mosquitoes homozygous for an autosomal recessive mutation at the red-eye (r) locus have a brick-red eye color in larvae, pupae, and young adults, in contrast to the almost black color of the wild eye. Mosquitoes homozygous for this mutant allele have levels of ommochrome precursors that are indistinguishable from the wild-type, but the low-mobility xanthommatin spot is ochre-brown in color rather than red-brown as in the wild-type. Mosquitoes with two different mutant alleles at the X-linked pink-eye locus (p, which confers a pink eye color, and pw, which confers a white eye phenotype in homozygotes or hemizygous males) have normal levels of ommochrome precursors but no detectable xanthommatins. Mosquitoes homozygous for both the r and p mutant alleles have apricot-colored eyes and show no detectable xanthommatins. Both the pink-eye and red-eye mutations appear to involve defects in the transport into or assembly of pigments in the membrane-bound pigment granules rather then defects in ommochrome synthesis.

Increased production of mitochondrial reactive oxygen species and reduced adult life span in an insecticide-resistant strain of Anopheles gambiae

PubMed Central

Otali, Dennis; Novak, Robert J.; Wan, Wen; Bu, Su; Moellering, Douglas R.; De Luca, Maria

2014-01-01

Control of the malaria vector An. gambiae is still largely obtained through chemical intervention using pyrethroids, such as permethrin. However, strains of An. gambiae that are resistant to the toxic effects of pyrethroids have become widespread in several endemic areas over the last decade. The objective of this study was to assess differences in five life-history traits (larval developmental time and the body weight, fecundity, hatch rate, and longevity of adult females) and energy metabolism between a strain of An. gambiae that is resistant to permethrin (RSP), due to knockdown resistance and enhanced metabolic detoxification, and a permethrin susceptible strain reared under laboratory conditions. We also quantified the expression levels of five antioxidant enzyme genes: GSTe3, CAT, GPXH1, SOD1, and SOD2. We found that the RSP strain had a longer developmental time than the susceptible strain. Additionally, RSP adult females had higher wet body weight and increased water and glycogen levels. Compared to permethrin susceptible females, RSP females displayed reduced metabolic rate and mitochondrial coupling efficiency and higher mitochondrial ROS production. Furthermore, despite higher levels of GSTe3 and CAT transcripts, RSP females had a shorter adult life span than susceptible females. Collectively, these results suggest that permethrin resistance alleles might affect energy metabolism, oxidative stress, and adult survival of An. gambiae. However, because the strains used in this study differ in their genetic backgrounds, the results need to be interpreted with caution and replicated in other strains in order to have significant implications for malaria transmission and vector control. PMID:24555527

Spatio-temporal genetic structure of Anopheles gambiae in the Northwestern Lake Victoria Basin, Uganda: implications for genetic control trials in malaria endemic regions.

PubMed

Lukindu, Martin; Bergey, Christina M; Wiltshire, Rachel M; Small, Scott T; Bourke, Brian P; Kayondo, Jonathan K; Besansky, Nora J

2018-04-16

Understanding population genetic structure in the malaria vector Anopheles gambiae (s.s.) is crucial to inform genetic control and manage insecticide resistance. Unfortunately, species characteristics such as high nucleotide diversity, large effective population size, recent range expansion, and high dispersal ability complicate the inference of genetic structure across its range in sub-Saharan Africa. The ocean, along with the Great Rift Valley, is one of the few recognized barriers to gene flow in this species, but the effect of inland lakes, which could be useful sites for initial testing of genetic control strategies, is relatively understudied. Here we examine Lake Victoria as a barrier between the Ugandan mainland and the Ssese Islands, which lie up to 60 km offshore. We use mitochondrial DNA (mtDNA) from populations sampled in 2002, 2012 and 2015, and perform Bayesian cluster analysis on mtDNA combined with microsatellite data previously generated from the same 2002 mosquito DNA samples. Hierarchical analysis of molecular variance and Bayesian clustering support significant differentiation between the mainland and lacustrine islands. In an mtDNA haplotype network constructed from this and previous data, haplotypes are shared even between localities separated by the Rift Valley, a result that more likely reflects retention of shared ancestral polymorphism than contemporary gene flow. The relative genetic isolation of An. gambiae on the Ssese Islands, their small size, level terrain and ease of access from the mainland, the relative simplicity of the vectorial system, and the prevalence of malaria, are all attributes that recommend these islands as possible sites for the testing of genetic control strategies.

Development of a DNA-Based Method for Distinguishing the Malaria Vectors, Anopheles gambiae From Anopheles arabiensis.

DTIC Science & Technology

1986-06-01

our preliminary studies hybridization with the Droso- phila actin probe required such low stringency conditions that the signal to noise ratio made...Balabacensis complex of Southeast Asia (Diptera: Culicidae). Genetica 57:81-86. (14) Mahon RJ and PM Miethke. 1982. Anopheles farauti No. 3, a hitherto un

Comparison of field-based xenodiagnosis and direct membrane feeding assays for evaluating host infectiousness to malaria vector Anopheles gambiae.

PubMed

Gouagna, Louis Clément; Yao, Frank; Yameogo, Bienvenue; Dabiré, Roch K; Ouédraogo, Jean-Bosco

2014-02-01

Several techniques are currently being used to study host infectiousness to mosquitoes, including the experimental possibility of laboratory reared mosquitoes acquiring infections through membrane feeders or directly on host skin. Here, the relative performance of the laboratory-based membrane feeding method (DMFA) and the field-based xenodiagnosis (XD) of malaria infectious hosts using wild Anopheles mosquitoes were compared. A cross-sectional survey involving a sample of 70 children (aged 3-12 years) living in a malaria endemic area in Western Burkina Faso, was carried out to measure their infectiousness to Anopheles mosquitoes using two approaches. The first approach used the xenodiagnostic procedure in which children were exposed to mosquito bites overnight, being sleeping individually in different sentinel huts from 6 pm to 6 am (4 nights per child). Anopheles sp that had acquired blood-meal on each child were subsequently collected early in the morning, and examined for Plasmodium falciparum oocyst infection on day 7 post-feeding. In the second approach, the infectiousness of the same children was estimated by whole-blood membrane feeding procedure using F0 An. gambiae s.l. that emerged from field-collected larvae cohorts. In the DMFA, 41.4% of the children successfully infected at least one mosquito with the mean oocyst prevalence of only 4.6±1.1% in the 2171 mosquitoes that were examined (mean oocyst intensity: 2.0±(std error of mean) 0.3 oocysts per infected midgut). Comparatively 78.6% of children yielded oocysts infection in mosquitoes during the XD approach (Chi square=20.11, df=1; p<0.001), with a mean rate of 19.6±2.0 in the 3752 wild caught mosquitoes (mean intensity: 3.93±0.2 oocysts per infected mosquito). The DMFA failed to reveal a portion (n=26) of infectious individuals that were sharply evidenced by the XD, particularly at low gametocyte densities or at levels that could not be detected by the classical microscopic examination of blood

Insecticide resistance in Bemisia tabaci Gennadius (Homoptera: Aleyrodidae) and Anopheles gambiae Giles (Diptera: Culicidae) could compromise the sustainability of malaria vector control strategies in West Africa.

PubMed

Gnankiné, Olivier; Bassolé, Imael H N; Chandre, Fabrice; Glitho, Isabelle; Akogbeto, Martin; Dabiré, Roch K; Martin, Thibaud

2013-10-01

Insecticides from the organophosphate (OP) and pyrethroid (PY) chemical families, have respectively, been in use for 50 and 30 years in West Africa, mainly against agricultural pests, but also against vectors of human disease. The selection pressure, with practically the same molecules year after year (mainly on cotton), has caused insecticide resistance in pest populations such as Bemisia tabaci, vector of harmful phytoviruses on vegetables. The evolution toward insecticide resistance in malaria vectors such as Anopheles gambiae sensus lato (s.l.) is probably related to the current use of these insecticides in agriculture. Thus, successful pest and vector control in West Africa requires an investigation of insect susceptibility, in relation to the identification of species and sub species, such as molecular forms or biotypes. Identification of knock down resistance (kdr) and acetylcholinesterase gene (Ace1) mutations modifying insecticide targets in individual insects and measure of enzymes activity typically involved in insecticide metabolism (oxidase, esterase and glutathion-S-transferase) are indispensable in understanding the mechanisms of resistance. Insecticide resistance is a good example in which genotype-phenotype links have been made successfully. Insecticides used in agriculture continue to select new resistant populations of B. tabaci that could be from different biotype vectors of plant viruses. As well, the evolution of insecticide resistance in An. gambiae threatens the management of malaria vectors in West Africa. It raises the question of priority in the use of insecticides in health and/or agriculture, and more generally, the question of sustainability of crop protection and vector control strategies in the region. Here, we review the susceptibility tests, biochemical and molecular assays data for B. tabaci, a major pest in cotton and vegetable crops, and An. gambiae, main vector of malaria. The data reviewed was collected in Benin and Burkina

Anopheles gambiae distribution and insecticide resistance in the cities of Douala and Yaoundé (Cameroon): influence of urban agriculture and pollution

PubMed Central

2011-01-01

Background Urban malaria is becoming a major health priority across Africa. A study was undertaken to assess the importance of urban pollution and agriculture practice on the distribution and susceptibility to insecticide of malaria vectors in the two main cities in Cameroon. Methods Anopheline larval breeding sites were surveyed and water samples analysed monthly from October 2009 to December 2010. Parameters analysed included turbidity, pH, temperature, conductivity, sulfates, phosphates, nitrates, nitrites, ammonia, aluminium, alkalinity, iron, potassium, manganese, magnesium, magnesium hardness and total hardness. Characteristics of water bodies in urban areas were compared to rural areas and between urban sites. The level of susceptibility of Anopheles gambiae to 4% DDT, 0.75% permethrin, 0.05% deltamethrin, 0.1% bendiocarb and 5% malathion were compared between mosquitoes collected from polluted, non polluted and cultivated areas. Results A total of 1,546 breeding sites, 690 in Yaoundé and 856 in Douala, were sampled in the course of the study. Almost all measured parameters had a concentration of 2- to 100-fold higher in urban compare to rural breeding sites. No resistance to malathion was detected, but bendiocarb resistance was present in Yaounde. Very low mortality rates were observed following DDT or permethrin exposure, associated with high kdr frequencies. Mosquitoes collected in cultivated areas, exhibited the highest resistant levels. There was little difference in insecticide resistance or kdr allele frequency in mosquitoes collected from polluted versus non-polluted sites. Conclusion The data confirm high selection pressure on mosquitoes originating from urban areas and suggest urban agriculture rather than pollution as the major factor driving resistance to insecticide. PMID:21651761

Insecticide susceptibility of natural populations of Anopheles coluzzii and Anopheles gambiae (sensu stricto) from Okyereko irrigation site, Ghana, West Africa.

PubMed

Chabi, Joseph; Baidoo, Philip K; Datsomor, Alex K; Okyere, Dora; Ablorde, Aikins; Iddrisu, Alidu; Wilson, Michael D; Dadzie, Samuel K; Jamet, Helen P; Diclaro, Joseph W

2016-03-31

The increasing spread of insecticide resistance in malaria vectors has been well documented across sub-Saharan Africa countries. The influence of irrigation on increasing vector resistance is poorly understood, and is critical to successful and ethical implementation of food security policies. This study investigated the insecticide resistance status of An. gambiae (s.l.) mosquitoes collected from the irrigated rice area of Okyereko, a village containing about 42 hectares of irrigated field within an irrigation project plan in the Central Region of Ghana. Large amounts of insecticides, herbicides and fertilizers are commonly used in the area to boost the annual production of the rice. Mosquito larvae were collected and adults were assayed from the F1 progeny. The resistance status, allele and genotype were characterized using WHO susceptibility testing and PCR methods respectively. The An. gambiae (s.l.) populations from Okyereko are highly resistant to DDT and pyrethroid insecticides, with possible involvement of metabolic mechanisms including the elevation of P450 and GST enzyme as well as P-gp activity. The population was mostly composed of An. coluzzii specimens (more than 96 %) with kdr and ace-1 frequencies of 0.9 and 0.2 %, respectively. This study brings additional information on insecticide resistance and the characterization of An. gambiae (s.l.) mosquitoes from Okyereko, which can be helpful in decision making for vector control programmes in the region.

Impact of PermaNet 3.0 on entomological indices in an area of pyrethroid resistant Anopheles gambiae in south-western Nigeria

PubMed Central

2014-01-01

Background PermaNet® 3.0 is an insecticide synergist-combination long-lasting insecticidal net designed to have increased efficacy against malaria vectors with metabolic resistance, even when combined with kdr. The current study reports on the impact of this improved tool on entomological indices in an area with pyrethroid-resistant malaria vectors in Nigeria. Methods Baseline entomological indices across eight villages in Remo North LGA of Ogun State provided the basis for selection of three villages (Ilara, Irolu and Ijesa) for comparing the efficacy of PermaNet® 3.0 (PN3.0), PermaNet® 2.0 (PN2.0) and untreated polyester nets as a control (UTC). In each case, nets were distributed to cover all sleeping spaces and were evaluated for insecticidal activity on a 3-monthly basis. Collection of mosquitoes was conducted monthly via window traps and indoor resting catches. The arithmetic means of mosquito catches per house, entomological inoculation rates before and during the intervention were compared as well as three other outcome parameters: the mean mosquito blood feeding rate, mean mortality and mean parity rates. Results Anopheles gambiae s.l. was the main malaria vector in the three villages, accounting for >98% of the Anopheles population and found in appreciable numbers for 6–7 months. Deltamethrin, permethrin and lambdacyhalothrin resistance were confirmed at Ilara, Irolu and Ijesa. The kdr mutation was the sole resistance mechanism at Ilara, whereas kdr plus P450-based metabolic mechanisms were detected at Irolu and Ijesa. Bioassays repeated on domestically used PN 2.0 and PN 3.0 showed persistent optimal (100%) bio-efficacy for both net types after the 3rd, 6th, 9th and 12th month following net distribution. The use of PN 3.0 significantly reduced mosquito densities with a ‘mass killing’ effect inside houses. Households with PN 3.0 also showed reduced blood feeding as well as lower mosquito parity and sporozoite rates compared to the PN 2.0 and the

A new tent trap for sampling exophagic and endophagic members of the Anopheles gambiae complex.

PubMed

Govella, Nicodemus J; Chaki, Prosper P; Geissbuhler, Yvonne; Kannady, Khadija; Okumu, Fredros; Charlwood, J Derek; Anderson, Robert A; Killeen, Gerry F

2009-07-14

Mosquito sampling methods are essential for monitoring and evaluating malaria vector control interventions. In urban Dar es Salaam, human landing catch (HLC) is the only method sufficiently sensitive for monitoring malaria-transmitting Anopheles. HLC is labour intensive, cumbersome, hazardous, and requires such intense supervision that is difficulty to sustain on large scales. Novel tent traps were developed as alternatives to HLC. The Furvela tent, designed in Mozambique, incorporates a CDC Light trap (LT) components, while two others from Ifakara, Tanzania (designs A and B) require no electricity or moving parts. Their sensitivity for sampling malaria vectors was compared with LT and HLC over a wide range of vector abundances in rural and urban settings in Tanzania, with endophagic and exophagic populations, respectively, using randomised Latin-square and cross- over experimental designs. The sensitivity of LTs was greater than HLC while the opposite was true of Ifakara tent traps (crude mean catch of An. gambiae sensu lato relative to HLC = 0.28, 0.65 and 1.30 for designs A, B and LT in a rural setting and 0.32 for design B in an urban setting). However, Ifakara B catches correlated far better to HLC (r2 = 0.73, P < 0.001) than any other method tested (r2 = 0.04, P = 0.426 and r2 = 0.19, P = 0.006 for Ifakara A and LTs respectively). Only Ifakara B in a rural setting with high vector density exhibited constant sampling efficiency relative to HLC. The relative sensitivity of Ifakara B increased as vector densities decreased in the urban setting and exceeded that of HLC at the lowest densities. None of the tent traps differed from HLC in terms of the proportions of parous mosquitoes (P >or= 0.849) or An. gambiae s.l. sibling species (P >or= 0.280) they sampled but both Ifakara A and B designs failed to reduce the proportion of blood-fed mosquitoes caught (Odds ratio [95% Confidence Interval] = 1.6 [1.2, 2.1] and 1.0 [0.8, 1.2], P = 0.002 and 0.998, respectively

Transcriptome of the adult female malaria mosquito vector Anopheles albimanus.

PubMed

Martínez-Barnetche, Jesús; Gómez-Barreto, Rosa E; Ovilla-Muñoz, Marbella; Téllez-Sosa, Juan; García López, David E; Dinglasan, Rhoel R; Ubaida Mohien, Ceereena; MacCallum, Robert M; Redmond, Seth N; Gibbons, John G; Rokas, Antonis; Machado, Carlos A; Cazares-Raga, Febe E; González-Cerón, Lilia; Hernández-Martínez, Salvador; Rodríguez López, Mario H

2012-05-30

Human Malaria is transmitted by mosquitoes of the genus Anopheles. Transmission is a complex phenomenon involving biological and environmental factors of humans, parasites and mosquitoes. Among more than 500 anopheline species, only a few species from different branches of the mosquito evolutionary tree transmit malaria, suggesting that their vectorial capacity has evolved independently. Anopheles albimanus (subgenus Nyssorhynchus) is an important malaria vector in the Americas. The divergence time between Anopheles gambiae, the main malaria vector in Africa, and the Neotropical vectors has been estimated to be 100 My. To better understand the biological basis of malaria transmission and to develop novel and effective means of vector control, there is a need to explore the mosquito biology beyond the An. gambiae complex. We sequenced the transcriptome of the An. albimanus adult female. By combining Sanger, 454 and Illumina sequences from cDNA libraries derived from the midgut, cuticular fat body, dorsal vessel, salivary gland and whole body, we generated a single, high-quality assembly containing 16,669 transcripts, 92% of which mapped to the An. darlingi genome and covered 90% of the core eukaryotic genome. Bidirectional comparisons between the An. gambiae, An. darlingi and An. albimanus predicted proteomes allowed the identification of 3,772 putative orthologs. More than half of the transcripts had a match to proteins in other insect vectors and had an InterPro annotation. We identified several protein families that may be relevant to the study of Plasmodium-mosquito interaction. An open source transcript annotation browser called GDAV (Genome-Delinked Annotation Viewer) was developed to facilitate public access to the data generated by this and future transcriptome projects. We have explored the adult female transcriptome of one important New World malaria vector, An. albimanus. We identified protein-coding transcripts involved in biological processes that may

Detection of knockdown resistance (kdr) mutations in Anopheles gambiae: a comparison of two new high-throughput assays with existing methods

PubMed Central

Bass, Chris; Nikou, Dimitra; Donnelly, Martin J; Williamson, Martin S; Ranson, Hilary; Ball, Amanda; Vontas, John; Field, Linda M

2007-01-01

Background Knockdown resistance (kdr) is a well-characterized mechanism of resistance to pyrethroid insecticides in many insect species and is caused by point mutations of the pyrethroid target site the para-type sodium channel. The presence of kdr mutations in Anopheles gambiae, the most important malaria vector in Africa, has been monitored using a variety of molecular techniques. However, there are few reports comparing the performance of these different assays. In this study, two new high-throughput assays were developed and compared with four established techniques. Methods Fluorescence-based assays based on 1) TaqMan probes and 2) high resolution melt (HRM) analysis were developed to detect kdr alleles in An. gambiae. Four previously reported techniques for kdr detection, Allele Specific Polymerase Chain Reaction (AS-PCR), Heated Oligonucleotide Ligation Assay (HOLA), Sequence Specific Oligonucleotide Probe – Enzyme-Linked ImmunoSorbent Assay (SSOP-ELISA) and PCR-Dot Blot were also optimized. The sensitivity and specificity of all six assays was then compared in a blind genotyping trial of 96 single insect samples that included a variety of kdr genotypes and African Anopheline species. The relative merits of each assay was assessed based on the performance in the genotyping trial, the length/difficulty of each protocol, cost (both capital outlay and consumable cost), and safety (requirement for hazardous chemicals). Results The real-time TaqMan assay was both the most sensitive (with the lowest number of failed reactions) and the most specific (with the lowest number of incorrect scores). Adapting the TaqMan assay to use a PCR machine and endpoint measurement with a fluorimeter showed a slight reduction in sensitivity and specificity. HRM initially gave promising results but was more sensitive to both DNA quality and quantity and consequently showed a higher rate of failure and incorrect scores. The sensitivity and specificity of AS-PCR, SSOP-ELISA, PCR Dot

Larval competition between An. coluzzii and An. gambiae in insectary and semi-field conditions in Burkina Faso.

PubMed

Gimonneau, Geoffrey; Brossette, Lou; Mamaï, Wadaka; Dabiré, Roch K; Simard, Frédéric

2014-02-01

Competition in mosquito larvae is common and different ecological context could change competitive advantage between species. Here, larval competition between the widely sympatric African malaria mosquitoes, Anopheles coluzzii and Anopheles gambiae were investigated in controlled insectary conditions using individuals from laboratory colonies and under ambient conditions using wild mosquitoes in a semi-field enclosure in western Burkina Faso. Larvae of both species were reared in trays at the same larval density and under the same feeding regimen in either single-species or mixed-species populations at varying species ratios reflecting 0%, 25%, 50% and 75% of competitor species. In the insectaries, where environmental variations are controlled, larvae of the An. coluzzii colony developed faster and with lower mortality than larvae of the An. gambiae colony (8.8±0.1 days and 21±3% mortality vs. 9.5±0.1 days and 32±3% mortality, respectively). Although there was no significant effect of competition on these phenotypic traits in any species, there was a significant trend for higher fitness of the An. coluzzii colony when competing with An. gambiae under laboratory conditions (i.e. lower development time and increased wing length at emergence, Cuzik's tests, P<0.05). In semi-field experiments, competition affected the life history traits of both species in a different way. Larvae of An. gambiae tended to reduce development time when in competition with An. coluzzii (Cuzick's test, P=0.002) with no impact either on mortality or size at emergence. On the other hand, An. coluzzii showed a significant trend for reduced larval mortality with increasing competition pressure (Cuzick's test, P=0.037) and production of smaller females when grown together with An. gambiae (Cuzick's test, P=0.002). Our results hence revealed that competitive interactions between larvae of the two species are context dependent. They further call for caution when exploring ecological processes

Potential use of neem leaf slurry as a sustainable dry season management strategy to control the malaria vector Anopheles gambiae (DIPTERA: CULICIDAE) in west African villages.

PubMed

Luong, Kyphuong; Dunkel, Florence V; Coulibaly, Keriba; Beckage, Nancy E

2012-11-01

Larval management of the malaria vector, Anopheles gambiae Giles s.s., has been successful in reducing disease transmission. However, pesticides are not affordable to farmers in remote villages in Mali, and in other material resource poor countries. Insect resistance to insecticides and nontarget toxicity pose additional problems. Neem (Azadirachta indica A. Juss) is a tree with many beneficial, insect bioactive compounds, such as azadirachtin. We tested the hypothesis that neem leaf slurry is a sustainable, natural product, anopheline larvicide. A field study conducted in Sanambele (Mali) in 2010 demonstrated neem leaf slurry can work with only the available tools and resources in the village. Laboratory bioassays were conducted with third instar An. gambiae and village methods were used to prepare the leaf slurry. Experimental concentration ranges were 1,061-21,224 mg/L pulverized neem leaves in distilled water. The 50 and 90% lethal concentrations at 72 h were 8,825 mg/L and 15,212 mg/L, respectively. LC concentrations were higher than for other parts of the neem tree when compared with previous published studies because leaf slurry preparation was simplified by omitting removal of fibrous plant tissue. Using storytelling as a medium of knowledge transfer, villagers combined available resources to manage anopheline larvae. Preparation of neem leaf slurries is a sustainable approach which allows villagers to proactively reduce mosquito larval density within their community as part of an integrated management system.

Structural and functional characterization of the contractile aorta and associated hemocytes of the mosquito Anopheles gambiae.

PubMed

Sigle, Leah T; Hillyer, Julián F

2018-06-22

The primary pump of the circulatory system of insects is a dorsal vessel that traverses the length of the insect. The anterior portion, located in the head, neck and thorax, is the aorta, and the posterior portion, located in the abdomen, is the heart. Here, we characterize the structure and function of the aorta and conical chamber of the mosquito, Anopheles gambiae The aorta begins in the head with an excurrent opening located above the dorsal pharyngeal plate and ends at the thoraco-abdominal junction where it joins the conical chamber of the heart. The aorta lacks ostia, and based on the diameter of the vessel as well as the density and helical orientation of muscle, consists of three regions: the anterior aorta, the bulbous chamber, and the posterior aorta. The aorta contracts in the anterograde direction, but these contractions are independent of heart contractions and do not play a major role in hemolymph propulsion. Intravital imaging of the venous channels, the first abdominal segment and the neck revealed that hemolymph only travels through the aorta in the anterograde direction, and does so only during periods of anterograde heart flow. Furthermore, hemolymph only enters the thoraco-abdominal ostia of the conical chamber when the heart contracts in the retrograde direction, propelling this hemolymph to the posterior of the body. Finally, very few hemocytes associate with the aorta, and unlike what is seen in the periostial regions of the heart, infection does not induce the aggregation of hemocytes on the aorta. © 2018. Published by The Company of Biologists Ltd.

Eater and draper are involved in the periostial haemocyte immune response in the mosquito Anopheles gambiae.

PubMed

Sigle, L T; Hillyer, J F

2018-03-09

Haemocytes respond to infection by phagocytosing pathogens, producing the enzymes that drive the phenoloxidase-based melanization cascade, secreting lytic factors, and producing other humoral proteins. A subset of haemocytes, called periostial haemocytes, aggregate on the surface of the heart of mosquitoes and kill pathogens in areas of high haemolymph flow. Periostial haemocytes are always present, but an infection induces the recruitment of additional haemocytes to these regions. Here, we tested whether members of the Nimrod gene family are involved in the periostial immune response of the African malaria mosquito, Anopheles gambiae. Using organismal manipulations, RNA interference (RNAi) and microscopy, we show that, following an infection with Escherichia coli, nimrod - the orthologue of Drosophila NimB2 - is not involved in periostial responses. At 4 h postinfection, however, RNAi-based knockdown of draper results in a marginal increase in the number of periostial haemocytes and a doubling of E. coli accumulation at the periostial regions. Finally, at 24 h postinfection, knockdown of eater decreases the number of periostial haemocytes and decreases the phagocytosis of E. coli on the surface of the heart. Phagocytosis of bacteria is more prevalent in the periostial regions of the mid abdominal segments, and knockdown of draper, nimrod or eater does not alter this distribution. Finally, knockdown of Nimrod family genes did not have a meaningful effect on the accumulation of melanin at the periostial regions. This study identifies roles for eater and draper in the functional integration of the mosquito immune and circulatory systems. © 2018 The Royal Entomological Society.

Composition of Anopheles mosquitoes, their blood-meal hosts, and Plasmodium falciparum infection rates in three islands with disparate bed net coverage in Lake Victoria, Kenya.

PubMed

Ogola, Edwin; Villinger, Jandouwe; Mabuka, Danspaid; Omondi, David; Orindi, Benedict; Mutunga, James; Owino, Vincent; Masiga, Daniel K

2017-09-08

Small islands serve as potential malaria reservoirs through which new infections might come to the mainland and may be important targets in malaria elimination efforts. This study investigated malaria vector species diversity, blood-meal hosts, Plasmodium infection rates, and long-lasting insecticidal net (LLIN) coverage on Mageta, Magare and Ngodhe Islands of Lake Victoria in western Kenya, a region where extensive vector control is implemented on the mainland. From trapping for six consecutive nights per month (November 2012 to March 2015) using CDC light traps, pyrethrum spray catches and backpack aspiration, 1868 Anopheles mosquitoes were collected. Based on their cytochrome oxidase I (COI) and intergenic spacer region PCR and sequencing, Anopheles gambiae s.l. (68.52%), Anopheles coustani (19.81%) and Anopheles funestus s.l. (11.67%) mosquitoes were differentiated. The mean abundance of Anopheles mosquitoes per building per trap was significantly higher (p < 0.001) in Mageta than in Magare and Ngodhe. Mageta was also the most populated island (n = 6487) with low LLIN coverage of 62.35% compared to Ngodhe (n = 484; 88.31%) and Magare (n = 250; 98.59%). Overall, 416 (22.27%) engorged Anopheles mosquitoes were analysed, of which 41 tested positive for Plasmodium falciparum infection by high-resolution melting (HRM) analysis of 18S rRNA and cytochrome b PCR products. Plasmodium falciparum infection rates were 10.00, 11.76, 0, and 18.75% among blood-fed An. gambiae s.s. (n = 320), Anopheles arabiensis (n = 51), An. funestus s.s. (n = 29), and An. coustani (n = 16), respectively. Based on HRM analysis of vertebrate cytochrome b, 16S rRNA and COI PCR products, humans (72.36%) were the prominent blood-meal hosts of malaria vectors, but 20.91% of blood-meals were from non-human vertebrate hosts. These findings demonstrate high Plasmodium infection rates among the primary malaria vectors An. gambiae s.s. and An. arabiensis, as well as in An. coustani

Infrared video tracking of Anopheles gambiae at insecticide-treated bed nets reveals rapid decisive impact after brief localised net contact

PubMed Central

Parker, Josephine E.A.; Angarita-Jaimes, Natalia; Abe, Mayumi; Towers, Catherine E.; Towers, David; McCall, Philip J.

2015-01-01

Long-lasting insecticidal bed nets (LLINs) protect humans from malaria transmission and are fundamental to malaria control worldwide, but little is known of how mosquitoes interact with nets. Elucidating LLIN mode of action is essential to maintain or improve efficacy, an urgent need as emerging insecticide resistance threatens their future. Tracking multiple free-flying Anopheles gambiae responding to human-occupied bed nets in a novel large-scale system, we characterised key behaviours and events. Four behavioural modes with different levels of net contact were defined: swooping, visiting, bouncing and resting. Approximately 75% of all activity occurred at the bed net roof where multiple brief contacts were focussed above the occupant’s torso. Total flight and net contact times were lower at LLINs than untreated nets but the essential character of the response was unaltered. LLINs did not repel mosquitoes but impacted rapidly: LLIN contact of less than 1 minute per mosquito during the first ten minutes reduced subsequent activity; after thirty minutes, activity at LLINs was negligible. Velocity measurements showed that mosquitoes detected nets, including unbaited untreated nets, prior to contact. This is the most complete characterisation of mosquito-LLIN interactions to date, and reveals many aspects of LLIN mode of action, important for developing the next generation of LLINs. PMID:26323965

A pre-intervention study of malaria vector abundance in Rio Muni, Equatorial Guinea: Their role in malaria transmission and the incidence of insecticide resistance alleles

PubMed Central

Ridl, Frances C; Bass, Chris; Torrez, Miguel; Govender, Dayanandan; Ramdeen, Varsha; Yellot, Lee; Edu, Amado Edjang; Schwabe, Christopher; Mohloai, Peter; Maharaj, Rajendra; Kleinschmidt, Immo

2008-01-01

Background Following the success of the malaria control intervention on the island of Bioko, malaria control by the use of indoor residual spraying (IRS) and long-lasting insecticide-treated nets (LLITN) was extended to Rio Muni, on the mainland part of Equatorial Guinea. This manuscript reports on the malaria vectors present and the incidence of insecticide resistant alleles prior to the onset of the programme. Methods Anopheles mosquitoes were captured daily using window traps at 30 sentinel sites in Rio Muni, from December 2006 to July 2007. The mosquitoes were identified to species and their sporozoite rates, knockdown resistance (kdr) and acetylcholinesterase (AChE) sensitivity measured, to define the role of vector species in malaria transmission and their potential susceptibility to insecticides. Results A total of 6,162 Anopheles mosquitoes were collected of which 4,808 were morphologically identified as Anopheles gambiae s.l., 120 Anopheles funestus, 1,069 Anopheles moucheti, and 165 Anopheles nili s.l.. Both M and S molecular forms of Anopheles gambiae s.s. and Anopheles melas were identified. Anopheles ovengensis and Anopheles carnevalei were the only two members of the An. nili group to be identified. Using the species-specific sporozoite rates and the average number of mosquitoes per night, the number of infective mosquitoes per trap per 100 nights for each species complex was calculated as a measure of transmission risk. Both kdr-w and kdr-e alleles were present in the S-form of An. gambiae s.s. (59% and 19% respectively) and at much lower frequencies in the M-form (9.7% and 1.8% respectively). The kdr-w and kdr-e alleles co-occurred in 103 S-form and 1 M-form specimens. No insensitive AChE was detected. Conclusion Anopheles gambiae s.s, a member of the Anopheles gambiae complex was shown to be the major vector in Rio Muni with the other three groups playing a relatively minor role in transmission. The demonstration of a high frequency of kdr alleles

Contemporary evolution of resistance at the major insecticide target site gene Ace-1 by mutation and copy number variation in the malaria mosquito Anopheles gambiae

PubMed Central

Weetman, David; Mitchell, Sara N; Wilding, Craig S; Birks, Daniel P; Yawson, Alexander E; Essandoh, John; Mawejje, Henry D; Djogbenou, Luc S; Steen, Keith; Rippon, Emily J; Clarkson, Christopher S; Field, Stuart G; Rigden, Daniel J; Donnelly, Martin J

2015-01-01

Functionally constrained genes are ideal insecticide targets because disruption is often fatal, and resistance mutations are typically costly. Synaptic acetylcholinesterase (AChE) is an essential neurotransmission enzyme targeted by insecticides used increasingly in malaria control. In Anopheles and Culex mosquitoes, a glycine–serine substitution at codon 119 of the Ace-1 gene confers both resistance and fitness costs, especially for 119S/S homozygotes. G119S in Anopheles gambiae from Accra (Ghana) is strongly associated with resistance, and, despite expectations of cost, resistant 119S alleles are increasing significantly in frequency. Sequencing of Accra females detected only a single Ace-1 119S haplotype, whereas 119G diversity was high overall but very low at non-synonymous sites, evidence of strong purifying selection driven by functional constraint. Flanking microsatellites showed reduced diversity, elevated linkage disequilibrium and high differentiation of 119S, relative to 119G homozygotes across up to two megabases of the genome. Yet these signals of selection were inconsistent and sometimes weak tens of kilobases from Ace-1. This unexpected finding is attributable to apparently ubiquitous amplification of 119S alleles as part of a large copy number variant (CNV) far exceeding the size of the Ace-1 gene, whereas 119G alleles were unduplicated. Ace-1 CNV was detectable in archived samples collected when the 119S allele was rare in Ghana. Multicopy amplification of resistant alleles has not been observed previously and is likely to underpin the recent increase in 119S frequency. The large CNV compromised localization of the strong selective sweep around Ace-1, emphasizing the need to integrate CNV analysis into genome scans for selection. PMID:25865270

Insecticide susceptibility of Anopheles mosquitoes changes in response to variations in the larval environment.

PubMed

Owusu, Henry F; Chitnis, Nakul; Müller, Pie

2017-06-16

Insecticide resistance threatens the success achieved through vector control in reducing the burden of malaria. An understanding of insecticide resistance mechanisms would help to develop novel tools and strategies to restore the efficacy of insecticides. Although we have substantially improved our understanding of the genetic basis of insecticide resistance over the last decade, we still know little of how environmental variations influence the mosquito phenotype. Here, we measured how variations in larval rearing conditions change the insecticide susceptibility phenotype of adult Anopheles mosquitoes. Anopheles gambiae and A. stephensi larvae were bred under different combinations of temperature, population density and nutrition, and the emerging adults were exposed to permethrin. Mosquitoes bred under different conditions showed considerable changes in mortality rates and body weight, with nutrition being the major factor. Weight is a strong predictor of insecticide susceptibility and bigger mosquitoes are more likely to survive insecticide treatment. The changes can be substantial, such that the same mosquito colony may be considered fully susceptible or highly resistant when judged by World Health Organization discriminatory concentrations. The results shown here emphasise the importance of the environmental background in developing insecticide resistance phenotypes, and caution for the interpretation of data generated by insecticide susceptibility assays.

The mode of action of spatial repellents and their impact on vectorial capacity of Anopheles gambiae sensu stricto.

PubMed

Ogoma, Sheila B; Ngonyani, Hassan; Simfukwe, Emmanuel T; Mseka, Antony; Moore, Jason; Maia, Marta F; Moore, Sarah J; Lorenz, Lena M

2014-01-01

Malaria vector control relies on toxicity of insecticides used in long lasting insecticide treated nets and indoor residual spraying. This is despite evidence that sub-lethal insecticides reduce human-vector contact and malaria transmission. The impact of sub-lethal insecticides on host seeking and blood feeding of mosquitoes was measured. Taxis boxes distinguished between repellency and attraction inhibition of mosquitoes by measuring response of mosquitoes towards or away from Transfluthrin coils and humans. Protective effective distance of coils and long-term effects on blood feeding were measured in the semi-field tunnel and in a Peet Grady chamber. Laboratory reared pyrethroid susceptible Anopheles gambiae sensu stricto mosquitoes were used. In the taxis boxes, a higher proportion of mosquitoes (67%-82%) were activated and flew towards the human in the presence of Transfluthrin coils. Coils did not hinder attraction of mosquitoes to the human. In the semi-field Tunnel, coils placed 0.3 m from the human reduced feeding by 86% (95% CI [0.66; 0.95]) when used as a "bubble" compared to 65% (95% CI [0.51; 0.76]) when used as a "point source". Mosquitoes exposed to coils inside a Peet Grady chamber were delayed from feeding normally for 12 hours but there was no effect on free flying and caged mosquitoes exposed in the semi-field tunnel. These findings indicate that airborne pyrethroids minimize human-vector contact through reduced and delayed blood feeding. This information is useful for the development of target product profiles of spatial repellent products that can be used to complement mainstream malaria vector control tools.

Field experiments of Anopheles gambiae attraction to local fruits/seedpods and flowering plants in Mali to optimize strategies for malaria vector control in Africa using attractive toxic sugar bait methods.

PubMed

Müller, Günter C; Beier, John C; Traore, Sekou F; Toure, Mahamoudou B; Traore, Mohamed M; Bah, Sekou; Doumbia, Seydou; Schlein, Yosef

2010-09-20

Based on recent studies in Israel demonstrating that attractive toxic sugar bait (ATSB) methods can be used to decimate local anopheline and culicine mosquito populations, an important consideration is whether the same methods can be adapted and improved to attract and kill malaria vectors in Africa. The ATSB approach uses fruit or flower scent as an attractant, sugar solution as a feeding stimulant, and an oral toxin. The ATSB solutions are either sprayed on vegetation or suspended in simple bait stations, and the mosquitoes ingesting the toxic solutions are killed. As such, this approach targets sugar-feeding female and male mosquitoes. This study examines the attractiveness of African malaria vectors to local fruits/seedpods and flowering plants, key biological elements of the ATSB approach for mosquito control. Three field experiments were conducted at sites in Mali. The attraction of Anopheles gambiae s.l. to 26 different local fruits and seedpods was determined at a site in the semi-arid Bandiagara District of Mali. Wire mesh glue traps with fruits/seedpods suspended on skewers inside were set along a seasonal lagoon. Seven replicates of each fruit/seedpod species were tested, with a water-soaked sponge and a sugar-soaked sponge as controls. The attraction of An. gambiae s.l. to 26 different types of flowering plants was determined at a site near Mopti in Mali. The flowering plants held in a water-filled buried container were tested using the same glue traps, with controls including water only and sugar solution. Six replicates of each selected plant type were tested on transects between rice paddies. Additional studies using CDC light traps were done to determine the relative densities and periodicity of An. gambiae s.l. attraction to branches of the most highly attractive flowering plant, branches without flowers, human odor, and candescent light. Of the 26 fruits and seedpods tested, 6 were attractive to An. gambiae s.l. females and males, respectively

MALDI-TOF MS identification of Anopheles gambiae Giles blood meal crushed on Whatman filter papers.

PubMed

Niare, Sirama; Almeras, Lionel; Tandina, Fatalmoudou; Yssouf, Amina; Bacar, Affane; Toilibou, Ali; Doumbo, Ogobara; Raoult, Didier; Parola, Philippe

2017-01-01

Identification of the source of mosquito blood meals is an important component for disease control and surveillance. Recently, matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF MS) profiling has emerged as an effective tool for mosquito blood meal identification, using the abdomens of freshly engorged mosquitoes. In the field, mosquito abdomens are crushed on Whatman filter papers to determine the host feeding patterns by identifying the origin of their blood meals. The aim of this study was to test whether crushing engorged mosquito abdomens on Whatman filter papers was compatible with MALDI-TOF MS for mosquito blood meal identification. Both laboratory reared and field collected mosquitoes were tested. Sixty Anopheles gambiae Giles were experimentally engorged on the blood of six distinct vertebrate hosts (human, sheep, rabbit, dog, chicken and rat). The engorged mosquito abdomens were crushed on Whatman filter papers for MALDI-TOF MS analysis. 150 Whatman filter papers, with mosquitoes engorged on cow and goat blood, were preserved. A total of 77 engorged mosquito abdomens collected in the Comoros Islands and crushed on Whatman filter papers were tested with MALDI-TOF MS. The MS profiles generated from mosquito engorged abdomens crushed on Whatman filter papers exhibited high reproducibility according to the original host blood. The blood meal host was correctly identified from mosquito abdomens crushed on Whatman filter papers by MALDI-TOF MS. The MS spectra obtained after storage were stable regardless of the room temperature and whether or not they were frozen. The MS profiles were reproducible for up to three months. For the Comoros samples, 70/77 quality MS spectra were obtained and matched with human blood spectra. This was confirmed by molecular tools. The results demonstrated that MALDI-TOF MS could identify mosquito blood meals from Whatman filter papers collected in the field during entomological surveys. The

Impact of agriculture on the selection of insecticide resistance in the malaria vector Anopheles gambiae: a multigenerational study in controlled conditions.

PubMed

Nkya, Theresia Estomih; Poupardin, Rodolphe; Laporte, Frederic; Akhouayri, Idir; Mosha, Franklin; Magesa, Stephen; Kisinza, William; David, Jean-Philippe

2014-10-16

Resistance of mosquitoes to insecticides is mainly attributed to their adaptation to vector control interventions. Although pesticides used in agriculture have been frequently mentioned as an additional force driving the selection of resistance, only a few studies were dedicated to validate this hypothesis and characterise the underlying mechanisms. While insecticide resistance is rising dramatically in Africa, deciphering how agriculture affects resistance is crucial for improving resistance management strategies. In this context, the multigenerational effect of agricultural pollutants on the selection of insecticide resistance was examined in Anopheles gambiae. An urban Tanzanian An. gambiae population displaying a low resistance level was used as a parental strain for a selection experiment across 20 generations. At each generation larvae were selected with a mixture containing pesticides and herbicides classically used in agriculture in Africa. The resistance levels of adults to deltamethrin, DDT and bendiocarb were compared between the selected and non-selected strains across the selection process together with the frequency of kdr mutations. A microarray approach was used for pinpointing transcription level variations selected by the agricultural pesticide mixture at the adult stage. A gradual increase of adult resistance to all insecticides was observed across the selection process. The frequency of the L1014S kdr mutation rose from 1.6% to 12.5% after 20 generations of selection. Microarray analysis identified 90 transcripts over-transcribed in the selected strain as compared to the parental and the non-selected strains. Genes encoding cuticle proteins, detoxification enzymes, proteins linked to neurotransmitter activity and transcription regulators were mainly affected. RT-qPCR transcription profiling of candidate genes across multiple generations supported their link with insecticide resistance. This study confirms the potency of agriculture in selecting

Studies of Anopheles gambiae s.l (Diptera: Culicidae) exhibiting different vectorial capacities in lymphatic filariasis transmission in the Gomoa district, Ghana.

PubMed

Amuzu, Hilaria; Wilson, Michael D; Boakye, Daniel A

2010-09-14

Two lymphatic filariasis endemic communities Mampong and Hwida in Ghana have been regularly monitored for impact on transmission after annual mass drug administration (MDA) with albendazole and ivermectin. After six MDAs even though the ABR for Mampong was 55883/person/year and that of Hwida was 2494/person/year, they both had ATPs of 15.21 infective larvae/person/year. Interestingly the human microfilaraemia levels had reduced significantly from 14% to 0% at Mampong and 12% to 3% at Hwida. In an attempt to understand this anomaly, we collected mosquitoes over a 5-month period using human landing catches to determine the species composition, the number of cibarial teeth, the lengths and widths of the cibarium and the cibarial dome of the vector populations. Out of 2553 mosquitoes caught at Mampong, 42.6% were An. gambiae s.l. All 280 identified further by PCR were An. gambiae s.s (275 M and 5 S molecular forms). At Hwida, 112 mosquitoes were obtained; 67 (59.8%) were An. gambiae s.l, comprised of 40 (59.7%) An. melas, 24 (35.8%) An. gambiae s.s (17 and 5 M and S molecular forms respectively) and 3 (4.5%) unidentified. The mean number of teeth for An. melas was 14.1 (median = 14, range = 12-15), An. gambiae s.s., 15.7 (median = 15, range = 13-19) M form 15.5 (median = 15 range = 13-19) and S form 16 (median = 16, range 15-17). The observed differences in teeth numbers were significantly different between An. melas and An. gambiae s.s (p = 0.004), and the M form (p = 0.032) and the S form (p = 0.002). In this study, An. gambiae s.s was the main vector at Mampong and was found to possess significantly more cibarial teeth than An. melas, the principal vector at Hwida. We postulate that the different impact observed after 6 MDAs may be due to An. gambiae s.s exhibiting 'facilitation' at Mampong and at Hwida An. melas the main vector exhibits 'limitation'. Thus it may be necessary to compliment MDA with vector control to achieve interruption of transmission in areas

Dissecting the mechanisms responsible for the multiple insecticide resistance phenotype in Anopheles gambiae s.s., M form, from Vallée du Kou, Burkina Faso

PubMed Central

Kwiatkowska, Rachel M.; Platt, Naomi; Poupardin, Rodolphe; Irving, Helen; Dabire, Roch K.; Mitchell, Sara; Jones, Christopher M.; Diabaté, Abdoulaye; Ranson, Hilary; Wondji, Charles S.

2013-01-01

With the exception of target site mutations, insecticide resistance mechanisms in the principle malaria vector Anopheles gambiae, remains largely uncharacterized in Burkina Faso. Here we detected high prevalence of resistance in Vallée du Kou (VK) to pyrethroids, DDT and dieldrin, moderate level for carbamates and full susceptibility to organophosphates. High frequencies of L1014F kdr (75%) and Rdl (87%) mutations were observed showing strong correlation with pyrethroids/DDT and dieldrin resistance. The frequency of ace1R mutation was low even in carbamate resistant mosquitoes. Microarray analysis identified genes significantly over-transcribed in VK. These include the cytochrome P450 genes, CYP6P3 and CYP6Z2, previously associated with pyrethroid resistance. Gene Ontology (GO) enrichment analysis suggested that elevated neurotransmitter activity is associated with resistance, with the over-transcription of target site resistance genes such as acetylcholinesterase and the GABA receptor. A rhodopsin receptor gene previously associated with pyrethroid resistance in Culex pipiens pallens was also over-transcribed in VK. This study highlights the complex network of mechanisms conferring multiple resistance in malaria vectors and such information should be taken into account when designing and implementing resistance control strategies. PMID:23380570

Evaluation of active ingredients and larvicidal activity of clove and cinnamon essential oils against Anopheles gambiae (sensu lato).

PubMed

Thomas, Adelina; Mazigo, Humphrey D; Manjurano, Alphaxard; Morona, Domenica; Kweka, Eliningaya J

2017-09-06

Mosquitoes are well-known vectors of many diseases including malaria and lymphatic filariasis. Uses of synthetic insecticides are associated with high toxicity, resistance, environmental pollution and limited alternative, effective synthetic insecticides. This study was undertaken to evaluate the larvicidal efficacy of clove and cinnamon essential oils against laboratory Anopheles gambiae (sensu stricto) and wild An. arabiensis larvae. The standard WHO guideline for larvicides evaluation was used, and the GC-MS machine was used for active compounds percentage composition analysis and structures identification. Probit regression analysis was used for LC 50 and LC 95 calculations while a t-test was used to test for significant differences between laboratory-reared and wild larvae populations in each concentration of plant extract. Mortality effect of clove and cinnamon essential oils against wild and laboratory-reared larvae had variations indicated by their LC 50 and LC 95 values. The mortality at different concentrations of cinnamon and clove post-exposure for wild and laboratory-reared larvae were dosage-dependent and were higher for cinnamon than for clove essential oils. The mortality effect following exposure to a blend of the two essential oils was higher for blends containing a greater proportion of cinnamon oil. In the chemical analysis of the active ingredients of cinnamon essential oil, the main chemical content was Eugenol, and the rarest was β-Linalool while for clove essential oil, the main chemical content was Eugenol and the rarest was Bicyclo. The essential oils showed a larvicidal effect which was concentration-dependent for both laboratory and wild collected larvae. The active ingredient compositions triggered different responses in mortality. Further research in small-scale should be conducted with concentrated extracted compounds.

The dynamics of pyrethroid resistance in Anopheles arabiensis from Zanzibar and an assessment of the underlying genetic basis

PubMed Central

2013-01-01

Background The emergence of pyrethroid resistance in the malaria vector, Anopheles arabiensis, threatens to undermine the considerable gains made towards eliminating malaria on Zanzibar. Previously, resistance was restricted to the island of Pemba while mosquitoes from Unguja, the larger of the two islands of Zanzibar, were susceptible. Here, we characterised the mechanism(s) responsible for resistance on Zanzibar using a combination of gene expression and target-site mutation assays. Methods WHO resistance bioassays were conducted using 1-5d old adult Anopheles gambiae s.l. collected between 2011 and 2013 across the archipelago. Synergist assays with the P450 inhibitor piperonyl-butoxide were performed in 2013. Members of the An. gambiae complex were PCR-identified and screened for target-site mutations (kdr and Ace-1). Gene expression in pyrethroid resistant An. arabiensis from Pemba was analysed using whole-genome microarrays. Results Pyrethroid resistance is now present across the entire Zanzibar archipelago. Survival to the pyrethroid lambda-cyhalothrin in bioassays conducted in 2013 was 23.5-54.3% on Unguja and 32.9-81.7% on Pemba. We present evidence that resistance is mediated, in part at least, by elevated P450 monoxygenases. Whole-genome microarray scans showed that the most enriched gene terms in resistant An. arabiensis from Pemba were associated with P450 activity and synergist assays with PBO completely restored susceptibility to pyrethroids in both islands. CYP4G16 was the most consistently over-expressed gene in resistant mosquitoes compared with two susceptible strains from Unguja and Dar es Salaam. Expression of this P450 is enriched in the abdomen and it is thought to play a role in hydrocarbon synthesis. Microarray and qPCR detected several additional genes putatively involved in this pathway enriched in the Pemba pyrethroid resistant population and we hypothesise that resistance may be, in part, related to alterations in the structure of the

An online tool for mapping insecticide resistance in major Anopheles vectors of human malaria parasites and review of resistance status for the Afrotropical region

PubMed Central

2014-01-01

Background Malaria control programmes across Africa and beyond are facing increasing insecticide resistance in the major anopheline vectors. In order to preserve or prolong the effectiveness of the main malaria vector interventions, up-to-date and easily accessible insecticide resistance data that are interpretable at operationally-relevant scales are critical. Herein we introduce and demonstrate the usefulness of an online mapping tool, IR Mapper. Methods A systematic search of published, peer-reviewed literature was performed and Anopheles insecticide susceptibility and resistance mechanisms data were extracted and added to a database after a two-level verification process. IR Mapper ( http://www.irmapper.com) was developed using the ArcGIS for JavaScript Application Programming Interface and ArcGIS Online platform for exploration and projection of these data. Results Literature searches yielded a total of 4,084 susceptibility data points for 1,505 populations, and 2,097 resistance mechanisms data points for 1,000 populations of Anopheles spp. tested via recommended WHO methods from 54 countries between 1954 and 2012. For the Afrotropical region, data were most abundant for populations of An. gambiae, and pyrethroids and DDT were more often used in susceptibility assays (51.1 and 26.8% of all reports, respectively) than carbamates and organophosphates. Between 2001 and 2012, there was a clear increase in prevalence and distribution of confirmed resistance of An. gambiae s.l. to pyrethroids (from 41 to 87% of the mosquito populations tested) and DDT (from 64 to 91%) throughout the Afrotropical region. Metabolic resistance mechanisms were detected in western and eastern African populations and the two kdr mutations (L1014S and L1014F) were widespread. For An. funestus s.l., relatively few populations were tested, although in 2010–2012 resistance was reported in 50% of 10 populations tested. Maps are provided to illustrate the use of IR Mapper and the distribution

The Gambia.

PubMed

1982-11-01

Attention in this discussion of the Republic of the Gambia is directed to the following: the people; geography; history; government and political conditions; economy; foreign relations; and relations between the US and Gambia. In 1980 Gambia's population was estimated to be 619,052; the 1980-81 annual growth rate estimate was 2.8%. The infant mortality rate is 217/1000; life expectancy is 32 years for men and 34 years for women. Almost 85% of the population live in rural areas. The principal ethnic groups are Mandinka, Fula, Wolof, Jola, and Serahuli, English is the official language, although most Gambians speak their native language. The Gambia, located on the bulge of West Africa, is low lying, with a maximum altitude of 73 meters above sea level. From what is known of its early history, the Gambia was once part of the Empire of Ghana and the Kingdom of the Songhais. By the 16th century, Portuguese slave traders and gold seekers had settled in the lower river area. During the late 17th and throughout the 18th century, England and France struggled continously for political and commercial supremacy in the regions of the Senegal and Gambia rivers. The Gambia achieved independence on February 18, 1965, as a constitutional monarchy within the British Commonwealth. Shortly thereafter the government proposed conversion from a monarchy to a republic with an elected president replacing the British monarch as chief of state. In 1970 a referendum was approved by the required majority, and the Gambia became a republic on April 24th. The government is divided into independent executive, legislative, and judicial branches. Executive power is vested in a president, popularly elected for a 5 year term. The Gambia is 1 of the few African countries with a genuine multiparty system. There is no army. The economy is almost totally agricultural. The small manfacturing sector accounts for about 5% of the gross domestic product. Few foreign investors have taken advantage of the

Near-infrared spectroscopy as a complementary age grading and species identification tool for African malaria vectors

USDA-ARS?s Scientific Manuscript database

Near-infrared spectroscopy (NIRS) was recently applied to age-grade and differentiate laboratory reared Anopheles gambiae sensu strico and Anopheles arabiensis sibling species of Anopheles gambiae sensu lato. In this study, we report further on the accuracy of this tool in simultaneously estimating ...

Feeding and resting behaviour of malaria vector, Anopheles arabiensis with reference to zooprophylaxis

PubMed Central

Mahande, Aneth; Mosha, Franklin; Mahande, Johnson; Kweka, Eliningaya

2007-01-01

Background The most important factor for effective zooprophylaxis in reducing malaria transmission is a predominant population of a strongly zoophilic mosquito, Anopheles arabiensis. The feeding preference behaviour of Anopheline mosquitoes was evaluated in odour-baited entry trap (OBET). Methods Mosquitoes were captured daily using odour-baited entry traps, light traps and hand catch both indoor and in pit traps. Experimental huts were used for release and recapture experiment. The mosquitoes collected were compared in species abundances. Results Anopheles arabiensis was found to account for over 99% of Anopheles species collected in the study area in Lower Moshi, Northern Tanzania. In experimental release/capture trials conducted at the Mabogini verandah huts, An. arabiensis was found to have higher exophilic tendency (80.7%) compared to Anopheles gambiae (59.7%) and Culex spp. (60.8%). OBET experiments conducted at Mabogini collected a total of 506 An. arabiensis in four different trials involving human, cattle, sheep, goat and pig. Odours from the cattle attracted 90.3% (243) compared to odours from human, which attracted 9.7% (26) with a significant difference at P = 0.005. Odours from sheep, goat and pig attracted 9.7%, 7.2% and 7.3%, respectively. Estimation of HBI in An. arabiensis collected from houses in three lower Moshi villages indicated lower ratios for mosquitoes collected from houses with cattle compared to those without cattles. HBI was also lower in mosquitoes collected outdoors (0.1–0.3) compared to indoor (0.4–0.9). Conclusion In discussing the results, reference has been made to observation of exophilic, zoophilic and feeding tendencies of An. arabiensis, which are conducive for zooprophylaxis. It is recommended that in areas with a predominant An. arabiensis population, cattle should be placed close to dwelling houses in order to maximize the effects of zooprophylaxis. Protective effects of human from malaria can further be enhanced by

Contributions of Anopheles larval control to malaria suppression in tropical Africa: review of achievements and potential.

PubMed

Walker, K; Lynch, M

2007-03-01

Malaria vector control targeting the larval stages of mosquitoes was applied successfully against many species of Anopheles (Diptera: Culicidae) in malarious countries until the mid-20th Century. Since the introduction of DDT in the 1940s and the associated development of indoor residual spraying (IRS), which usually has a more powerful impact than larval control on vectorial capacity, the focus of malaria prevention programmes has shifted to the control of adult vectors. In the Afrotropical Region, where malaria is transmitted mainly by Anopheles funestus Giles and members of the Anopheles gambiae Giles complex, gaps in information on larval ecology and the ability of An. gambiae sensu lato to exploit a wide variety of larval habitats have discouraged efforts to develop and implement larval control strategies. Opportunities to complement adulticiding with other components of integrated vector management, along with concerns about insecticide resistance, environmental impacts, rising costs of IRS and logistical constraints, have stimulated renewed interest in larval control of malaria vectors. Techniques include environmental management, involving the temporary or permanent removal of anopheline larval habitats, as well as larviciding with chemical or biological agents. This present review covers large-scale trials of anopheline larval control methods, focusing on field studies in Africa conducted within the past 15 years. Although such studies are limited in number and scope, their results suggest that targeting larvae, particularly in human-made habitats, can significantly reduce malaria transmission in appropriate settings. These approaches are especially suitable for urban areas, where larval habitats are limited, particularly when applied in conjunction with IRS and other adulticidal measures, such as the use of insecticide treated bednets.

Contemporary evolution of resistance at the major insecticide target site gene Ace-1 by mutation and copy number variation in the malaria mosquito Anopheles gambiae.

PubMed

Weetman, David; Mitchell, Sara N; Wilding, Craig S; Birks, Daniel P; Yawson, Alexander E; Essandoh, John; Mawejje, Henry D; Djogbenou, Luc S; Steen, Keith; Rippon, Emily J; Clarkson, Christopher S; Field, Stuart G; Rigden, Daniel J; Donnelly, Martin J

2015-06-01

Functionally constrained genes are ideal insecticide targets because disruption is often fatal, and resistance mutations are typically costly. Synaptic acetylcholinesterase (AChE) is an essential neurotransmission enzyme targeted by insecticides used increasingly in malaria control. In Anopheles and Culex mosquitoes, a glycine-serine substitution at codon 119 of the Ace-1 gene confers both resistance and fitness costs, especially for 119S/S homozygotes. G119S in Anopheles gambiae from Accra (Ghana) is strongly associated with resistance, and, despite expectations of cost, resistant 119S alleles are increasing significantly in frequency. Sequencing of Accra females detected only a single Ace-1 119S haplotype, whereas 119G diversity was high overall but very low at non-synonymous sites, evidence of strong purifying selection driven by functional constraint. Flanking microsatellites showed reduced diversity, elevated linkage disequilibrium and high differentiation of 119S, relative to 119G homozygotes across up to two megabases of the genome. Yet these signals of selection were inconsistent and sometimes weak tens of kilobases from Ace-1. This unexpected finding is attributable to apparently ubiquitous amplification of 119S alleles as part of a large copy number variant (CNV) far exceeding the size of the Ace-1 gene, whereas 119G alleles were unduplicated. Ace-1 CNV was detectable in archived samples collected when the 119S allele was rare in Ghana. Multicopy amplification of resistant alleles has not been observed previously and is likely to underpin the recent increase in 119S frequency. The large CNV compromised localization of the strong selective sweep around Ace-1, emphasizing the need to integrate CNV analysis into genome scans for selection. © 2015 The Authors. Molecular Ecology published by John Wiley & Sons Ltd.

The larvicidal effects of black pepper (Piper nigrum L.) and piperine against insecticide resistant and susceptible strains of Anopheles malaria vector mosquitoes.

PubMed

Samuel, Michael; Oliver, Shüné V; Coetzee, Maureen; Brooke, Basil D

2016-04-26

Insecticide resistance carries the potential to undermine the efficacy of insecticide based malaria vector control strategies. Therefore, there is an urgent need for new insecticidal compounds. Black pepper (dried fruit from the vine, Piper nigrum), used as a food additive and spice, and its principal alkaloid piperine, have previously been shown to have larvicidal properties. The aim of this study was to investigate the larvicidal effects of ground black pepper and piperine against third and fourth instar Anopheles larvae drawn from several laboratory-reared insecticide resistant and susceptible strains of Anopheles arabiensis, An. coluzzii, An. gambiae, An. quadriannulatus and An. funestus. Larvae were fed with mixtures of standard larval food and either ground black pepper or piperine in different proportions. Mortality was recorded 24 h after black pepper and 48 h after piperine were applied to the larval bowls. Black pepper and piperine mixtures caused high mortality in the An. gambiae complex strains, with black pepper proving significantly more toxic than piperine. The An. funestus strains were substantially less sensitive to black pepper and piperine which may reflect a marked difference in the feeding habits of this species compared to that of the Gambiae complex or a difference in food metabolism as a consequence of differences in breeding habitat between species. Insecticide resistant and susceptible strains by species proved equally susceptible to black pepper and piperine. It is concluded that black pepper shows potential as a larvicide for the control of certain malaria vector species.

Wild Anopheles funestus mosquito genotypes are permissive for infection with the rodent malaria parasite, Plasmodium berghei.

PubMed

Xu, Jiannong; Hillyer, Julián F; Coulibaly, Boubacar; Sacko, Madjou; Dao, Adama; Niaré, Oumou; Riehle, Michelle M; Traoré, Sekou F; Vernick, Kenneth D

2013-01-01

Malaria parasites undergo complex developmental transitions within the mosquito vector. A commonly used laboratory model for studies of mosquito-malaria interaction is the rodent parasite, P. berghei. Anopheles funestus is a major malaria vector in sub-Saharan Africa but has received less attention than the sympatric species, Anopheles gambiae. The imminent completion of the A. funestus genome sequence will provide currently lacking molecular tools to describe malaria parasite interactions in this mosquito, but previous reports suggested that A. funestus is not permissive for P. berghei development. An A. funestus population was generated in the laboratory by capturing female wild mosquitoes in Mali, allowing them to oviposit, and rearing the eggs to adults. These F1 progeny of wild mosquitoes were allowed to feed on mice infected with a fluorescent P. berghei strain. Fluorescence microscopy was used to track parasite development inside the mosquito, salivary gland sporozoites were tested for infectivity to mice, and parasite development in A. funestus was compared to A. gambiae. P. berghei oocysts were detectable on A. funestus midguts by 7 days post-infection. By 18-20 days post-infection, sporozoites had invaded the median and distal lateral lobes of the salivary glands, and hemocoel sporozoites were observed in the hemolymph. Mosquitoes were capable of infecting mice via bite, demonstrating that A. funestus supports the complete life cycle of P. berghei. In a random sample of wild mosquito genotypes, A. funestus prevalence of infection and the characteristics of parasite development were similar to that observed in A. gambiae-P. berghei infections. The data presented in this study establish an experimental laboratory model for Plasmodium infection of A. funestus, an important vector of human malaria. Studying A. funestus-Plasmodium interactions is now feasible in a laboratory setting. This information lays the groundwork for exploitation of the awaited genome

Ability of TEP1 in intestinal flora to modulate natural resistance of Anopheles dirus.

PubMed

Wang, Yanyan; Wang, Ying; Zhang, Jingru; Xu, Wenyue; Zhang, Jian; Huang, Fu Sheng

2013-08-01

Blocking transmission of malaria is a reliable way to control and eliminate infection. However, in-depth knowledge of the interaction between Plasmodium and mosquito is needed. Studies suggest that innate immunity is the main mechanism inhibiting development of malaria parasites in the mosquito. Recent studies have found that use of antibiotics that inhibit the mosquito gut flora can reduce the immune response of Anopheles gambiae, thereby contributing to the development of malaria parasites. In our study, we used the non susceptible model of Anopheles dirus-Plasmodium yoelii to explore the effect of Anopheles intestinal flora on the natural resistance of A. dirus to P. yoelii. We found that in mosquitoes infected with Plasmodium, the intestinal flora can regulate expression of thioester-containing protein (TEP1) via an RNAi gene-silencing approach. Our results suggest that in the absence of TEP1, the natural microbiota cannot suppress the development of P. yoelii in A. dirus. This suggests that AdTEP1 plays an important role in the resistance of A. dirus to P. yoelii. The intestinal flora may modulate the development of P. yoelii in A. dirus by regulating TEP1 expression. Copyright © 2013 Elsevier Inc. All rights reserved.

Quantification of the efficiency of treatment of Anopheles gambiae breeding sites with petroleum products by local communities in areas of insecticide resistance in the Republic of Benin

PubMed Central

Djouaka, Rousseau F; Bakare, Adekunle A; Bankole, Honore S; Doannio, Julien MC MC; Kossou, Hortense; Akogbeto, Martin C

2007-01-01

Background The emergence of Anopheles populations capable of withstanding lethal doses of insecticides has weakened the efficacy of most insecticide based strategies of vector control and, has highlighted the need for developing new insecticidal molecules or, improving the efficacy of existing insecticides or abandoning those to which resistance has emerged. The use of petroleum products (PP) against mosquito larvae had an immense success during early programmes of malaria control, but these compounds were abandoned and replaced in the 1950s by synthetic insecticides probably because of the high performances given by these new products. In the current context of vector resistance, it is important to elucidate the empirical use of PP by quantifying their efficiencies on resistant strains of Anopheles. Methods Larvae of Anopheles Ladji a local resistant strain were exposed to increasing concentrations of various PP (kerosene, petrol and engine oils) for 24 hours and the lethal activities recorded. The highest concentration (HiC) having no lethal activity (also referred as the NOEL or no effect level) and the lowest concentration (LoC100) yielding 100% mortality were rated for each PP on the Ladji strain. Prior to laboratory analysis, KAP studies were conducted in three traditional communities were insecticide resistance is clearly established to confirm the use of PP against mosquitoes. Results Laboratory analysis of petrol, kerosene and engine oils, clearly established their lethal activities on resistant strains of Anopheles larvae. Contrary to existing references, this research revealed that exposed larvae of Anopheles were mostly killed by direct contact toxicity and not by suffocation as indicated in some earlier reports. Conclusion This research could serve as scientific basis to backup the empirical utilisation of PP on mosquito larvae and to envisage possibilities of using PP in some traditional settings where Anopheles have developed resistance to currently

Anopheles gambiae distribution and insecticide resistance in the cities of Douala and Yaoundé (Cameroon): influence of urban agriculture and pollution.

PubMed

Antonio-Nkondjio, Christophe; Fossog, Billy Tene; Ndo, Cyrille; Djantio, Benjamin Menze; Togouet, Serge Zebaze; Awono-Ambene, Parfait; Costantini, Carlo; Wondji, Charles S; Ranson, Hilary

2011-06-08

Urban malaria is becoming a major health priority across Africa. A study was undertaken to assess the importance of urban pollution and agriculture practice on the distribution and susceptibility to insecticide of malaria vectors in the two main cities in Cameroon. Anopheline larval breeding sites were surveyed and water samples analysed monthly from October 2009 to December 2010. Parameters analysed included turbidity, pH, temperature, conductivity, sulfates, phosphates, nitrates, nitrites, ammonia, aluminium, alkalinity, iron, potassium, manganese, magnesium, magnesium hardness and total hardness. Characteristics of water bodies in urban areas were compared to rural areas and between urban sites. The level of susceptibility of Anopheles gambiae to 4% DDT, 0.75% permethrin, 0.05% deltamethrin, 0.1% bendiocarb and 5% malathion were compared between mosquitoes collected from polluted, non polluted and cultivated areas. A total of 1,546 breeding sites, 690 in Yaoundé and 856 in Douala, were sampled in the course of the study. Almost all measured parameters had a concentration of 2- to 100-fold higher in urban compare to rural breeding sites. No resistance to malathion was detected, but bendiocarb resistance was present in Yaounde. Very low mortality rates were observed following DDT or permethrin exposure, associated with high kdr frequencies. Mosquitoes collected in cultivated areas, exhibited the highest resistant levels. There was little difference in insecticide resistance or kdr allele frequency in mosquitoes collected from polluted versus non-polluted sites. The data confirm high selection pressure on mosquitoes originating from urban areas and suggest urban agriculture rather than pollution as the major factor driving resistance to insecticide.

The ace-1 Locus Is Amplified in All Resistant Anopheles gambiae Mosquitoes: Fitness Consequences of Homogeneous and Heterogeneous Duplications

PubMed Central

Djogbénou, Luc S.; Berthomieu, Arnaud; Makoundou, Patrick; Baba-Moussa, Lamine S.; Fiston-Lavier, Anna-Sophie; Belkhir, Khalid; Labbé, Pierrick; Weill, Mylène

2016-01-01

Gene copy-number variations are widespread in natural populations, but investigating their phenotypic consequences requires contemporary duplications under selection. Such duplications have been found at the ace-1 locus (encoding the organophosphate and carbamate insecticides’ target) in the mosquito Anopheles gambiae (the major malaria vector); recent studies have revealed their intriguing complexity, consistent with the involvement of various numbers and types (susceptible or resistant to insecticide) of copies. We used an integrative approach, from genome to phenotype level, to investigate the influence of duplication architecture and gene-dosage on mosquito fitness. We found that both heterogeneous (i.e., one susceptible and one resistant ace-1 copy) and homogeneous (i.e., identical resistant copies) duplications segregated in field populations. The number of copies in homogeneous duplications was variable and positively correlated with acetylcholinesterase activity and resistance level. Determining the genomic structure of the duplicated region revealed that, in both types of duplication, ace-1 and 11 other genes formed tandem 203kb amplicons. We developed a diagnostic test for duplications, which showed that ace-1 was amplified in all 173 resistant mosquitoes analyzed (field-collected in several African countries), in heterogeneous or homogeneous duplications. Each type was associated with different fitness trade-offs: heterogeneous duplications conferred an intermediate phenotype (lower resistance and fitness costs), whereas homogeneous duplications tended to increase both resistance and fitness cost, in a complex manner. The type of duplication selected seemed thus to depend on the intensity and distribution of selection pressures. This versatility of trade-offs available through gene duplication highlights the importance of large mutation events in adaptation to environmental variation. This impressive adaptability could have a major impact on vector

Mitochondrial genomes of Anopheles arabiensis, An.gambiae and An.coluzzii show no clear species division

USDA-ARS?s Scientific Manuscript database

Here we report the complete mitochondrial sequences of 70 individual field collected mosquito specimens from throughout Sub-Saharan Africa. We generated this dataset to identify species specific markers for the following Anopheles species and chromosomal forms: An.arabiensis, An.coluzzii (The Forest...

Temporal and micro-spatial heterogeneity in the distribution of Anopheles vectors of malaria along the Kenyan coast

PubMed Central

2013-01-01

Background The distribution of anopheline mosquitoes is determined by temporally dynamic environmental and human-associated variables, operating over a range of spatial scales. Macro-spatial short-term trends are driven predominantly by prior (lagged) seasonal changes in climate, which regulate the abundance of suitable aquatic larval habitats. Micro-spatial distribution is determined by the location of these habitats, proximity and abundance of available human bloodmeals and prevailing micro-climatic conditions. The challenge of analysing—in a single coherent statistical framework—the lagged and distributed effect of seasonal climate changes simultaneously with the effects of an underlying hierarchy of spatial factors has hitherto not been addressed. Methods Data on Anopheles gambiae sensu stricto and A. funestus collected from households in Kilifi district, Kenya, were analysed using polynomial distributed lag generalized linear mixed models (PDL GLMMs). Results Anopheline density was positively and significantly associated with amount of rainfall between 4 to 47 days, negatively and significantly associated with maximum daily temperature between 5 and 35 days, and positively and significantly associated with maximum daily temperature between 29 and 48 days in the past (depending on Anopheles species). Multiple-occupancy households harboured greater mosquito numbers than single-occupancy households. A significant degree of mosquito clustering within households was identified. Conclusions The PDL GLMMs developed here represent a generalizable framework for analysing hierarchically-structured data in combination with explanatory variables which elicit lagged effects. The framework is a valuable tool for facilitating detailed understanding of determinants of the spatio-temporal distribution of Anopheles. Such understanding facilitates delivery of targeted, cost-effective and, in certain circumstances, preventative antivectorial interventions against malaria

Habitat characterization and spatial distribution of Anopheles sp. mosquito larvae in Dar es Salaam (Tanzania) during an extended dry period

PubMed Central

Sattler, Michael A; Mtasiwa, Deo; Kiama, Michael; Premji, Zul; Tanner, Marcel; Killeen, Gerry F; Lengeler, Christian

2005-01-01

Introduction By 2030, more than 50% of the African population will live in urban areas. Controlling malaria reduces the disease burden and further improves economic development. As a complement to treated nets and prompt access to treatment, measures targeted against the larval stage of Anopheles sp. mosquitoes are a promising strategy for urban areas. However, a precise knowledge of the geographic location and potentially of ecological characteristics of breeding sites is of major importance for such interventions. Methods In total 151 km2 of central Dar es Salaam, the biggest city of Tanzania, were systematically searched for open mosquito breeding sites. Ecologic parameters, mosquito larvae density and geographic location were recorded for each site. Logistic regression analysis was used to determine the key ecological factors explaining the different densities of mosquito larvae. Results A total of 405 potential open breeding sites were examined. Large drains, swamps and puddles were associated with no or low Anopheles sp. larvae density. The probability of Anopheles sp. larvae to be present was reduced when water was identified as "turbid". Small breeding sites were more commonly colonized by Anopheles sp. larvae. Further, Anopheles gambiae s.l. larvae were found in highly organically polluted habitats. Conclusions Clear ecological characteristics of the breeding requirements of Anopheles sp. larvae could not be identified in this setting. Hence, every stagnant open water body, including very polluted ones, have to be considered as potential malaria vector breeding sites. PMID:15649333

Physiological correlates of ecological divergence along an urbanization gradient: differential tolerance to ammonia among molecular forms of the malaria mosquito Anopheles gambiae

PubMed Central

2013-01-01

Background Limitations in the ability of organisms to tolerate environmental stressors affect their fundamental ecological niche and constrain their distribution to specific habitats. Evolution of tolerance, therefore, can engender ecological niche dynamics. Forest populations of the afro-tropical malaria mosquito Anopheles gambiae have been shown to adapt to historically unsuitable larval habitats polluted with decaying organic matter that are found in densely populated urban agglomerates of Cameroon. This process has resulted in niche expansion from rural to urban environments that is associated with cryptic speciation and ecological divergence of two evolutionarily significant units within this taxon, the molecular forms M and S, among which reproductive isolation is significant but still incomplete. Habitat segregation between the two forms results in a mosaic distribution of clinally parapatric patches, with the M form predominating in the centre of urban agglomerates and the S form in the surrounding rural localities. We hypothesized that development of tolerance to nitrogenous pollutants derived from the decomposition of organic matter, among which ammonia is the most toxic to aquatic organisms, may affect this pattern of distribution and process of niche expansion by the M form. Results Acute toxicity bioassays indicated that populations of the two molecular forms occurring at the extremes of an urbanization gradient in Yaounde, the capital of Cameroon, differed in their response to ammonia. The regression lines best describing the dose-mortality profile differed in the scale of the explanatory variable (ammonia concentration log-transformed for the S form and linear for the M form), and in slope (steeper for the S form and shallower for the M form). These features reflected differences in the frequency distribution of individual tolerance thresholds in the two populations as assessed by probit analysis, with the M form exhibiting a greater mean and

Survival Value and Sugar Access of Four East African Plant Species Attractive to a Laboratory Strain of Sympatric Anopheles gambiae (Diptera: Culicidae)

PubMed Central

Nikbakhtzadeh, M. R.; Terbot, J. W.; Foster, W. A.

2016-01-01

Mosquitoes derive energy from plant sugar, thereby promoting survival and reproduction. Its survival value to females plays a key role in the vectorial capacity of mosquito populations. Previous olfactometry assays of responsiveness demonstrated that Senna didymobotrya Fresenius, Parthenium hysterophorus, L. Senna occidentalis, (L) and Lantana camara L were among the most attractive plants for the Mbita strain of Anopheles gambiae s.s. Giles in eastern Africa. Here, we provide experimental evidence that three of these four species also provide varying but substantial amounts of sugar for mosquito survival, whereas a fourth does not. Rank order of survival of both sexes of mosquitoes housed with these plants was as follows: S. didymobotrya was highest, followed by S. occidentalis and L. camara, whereas survival on P. hysterophorus was only slightly better than on only water. A positive control group, housed with 10% sucrose, survived well but fell significantly short of those with S. didymobotrya. A causal connection between survival and sugar availability was established by exposing mosquitoes to plants overnight, and then testing them for the presence and amount of undigested fructose. Fructose positivity was most frequent in those exposed to L. camara, whereas greatest amounts of fructose were obtained from S. occidentalis and S. didymobotrya. Parthenium hysterophorus scored lowest in both categories. We conclude that attractiveness and sugar availability are often, but not always, concordant. It remains unclear why P. hysterophorus should be attractive if it offers little sugar and does not prolong survival. Furthermore, the cause behind the superior survival benefit of S. didymobotrya, compared with 10% sucrose, is unknown. PMID:27247348

Development and survival of Anopheles gambiae eggs in drying soil: influence of the rate of drying, egg age, and soil type.

PubMed

Shililu, J I; Grueber, W B; Mbogo, C M; Githure, J I; Riddiford, L M; Beier, J C

2004-09-01

Little is known about the contribution made by the egg stage of African malaria vectors to the rapid rise in adult populations following the onset of seasonal rains. To examine this issue, we evaluated the viability of Anopheles gambiae eggs in drying soil in the laboratory. Survival data were collected from field-caught mosquitoes kept in sandy loam soil and laboratory-reared colonies kept in sandy loam soil and black cotton soil. Under high, medium, and low soil-moisture regimes, egg viability declined sharply with increased duration of drying. Eggs remained viable in drying sandy loam soil for 1, 5, and 10 days, but not after 15 or 20 days. The most dramatic decline in hatching success occurred between drying days 1 (78-83% hatch) and 5 (20-23% hatch). In contrast, eggs reared in high-moisture black cotton soil remained viable for up to 15 days. Furthermore, after 5 drying days, high-, medium-, and low-moisture soils averaged 59, 47, and 31% hatching success, respectively. We recovered unhatched eggs from sandy loam soils to examine the developmental status of the embryos. A majority of the unhatched eggs that were recovered from days 15 and 20 in sandy loam soils contained fully developed late-stage embryos. Thus, unhatched eggs completed embryonic development but probably died before receiving an appropriate hatching stimulus. Our results suggest that the absolute moisture content of the soil does not alone determine hatching success of anopheline eggs. Rather, soil moisture, together with the rate of drying, physiological factors associated with the age of the egg, and the type of soil in which the egg rests likely influence survival.

IgG responses to the gSG6-P1 salivary peptide for evaluating human exposure to Anopheles bites in urban areas of Dakar region, Sénégal

PubMed Central

2012-01-01

Background Urban malaria can be a serious public health problem in Africa. Human-landing catches of mosquitoes, a standard entomological method to assess human exposure to malaria vector bites, can lack sensitivity in areas where exposure is low. A simple and highly sensitive tool could be a complementary indicator for evaluating malaria exposure in such epidemiological contexts. The human antibody response to the specific Anopheles gSG6-P1 salivary peptide have been described as an adequate tool biomarker for a reliable assessment of human exposure level to Anopheles bites. The aim of this study was to use this biomarker to evaluate the human exposure to Anopheles mosquito bites in urban settings of Dakar (Senegal), one of the largest cities in West Africa, where Anopheles biting rates and malaria transmission are supposed to be low. Methods One cross-sectional study concerning 1,010 (505 households) children (n = 505) and adults (n = 505) living in 16 districts of downtown Dakar and its suburbs was performed from October to December 2008. The IgG responses to gSG6-P1 peptide have been assessed and compared to entomological data obtained in or near the same district. Results Considerable individual variations in anti-gSG6-P1 IgG levels were observed between and within districts. In spite of this individual heterogeneity, the median level of specific IgG and the percentage of immune responders differed significantly between districts. A positive and significant association was observed between the exposure levels to Anopheles gambiae bites, estimated by classical entomological methods, and the median IgG levels or the percentage of immune responders measuring the contact between human populations and Anopheles mosquitoes. Interestingly, immunological parameters seemed to better discriminate the exposure level to Anopheles bites between different exposure groups of districts. Conclusions Specific human IgG responses to gSG6-P1 peptide biomarker represent, at the

Effect of naturally-occurring Wolbachia in Anopheles gambiae s.l. mosquitoes from Mali on Plasmodium falciparum malaria transmission

USDA-ARS?s Scientific Manuscript database

A novel naturally-occurring Wolbachia strain was identified in mosquitoes of the A. gambiae complex collected in the Malian villages of Dangassa and Kenieroba (wAnga-Mali). Phylogenetic analysis of the nucleotide sequence of two 16S rRNA regions showed that wAnga-Mali clusters with Wolbachia strains...

Life-table analysis of Anopheles arabiensis in western Kenya highlands: effects of land covers on larval and adult survivorship.

PubMed

Afrane, Yaw A; Zhou, Goufa; Lawson, Bernard W; Githeko, Andrew K; Yan, Guiyun

2007-10-01

In high-elevation areas in western Kenya, the abundance of Anopheles arabiensis is either very low or absent. The western Kenya highlands (an area with an elevation > 1,500m above sea level) have also been experiencing extensive deforestation, and deforestation has been suggested as one of the important factors that facilitate malaria transmission in the highlands. This study investigated whether climate conditions in the western Kenya highlands (Kakamega, elevation 1,500 m above sea level) were permissive to the development and survival of An. arabiensis and whether deforestation promoted An. arabiensis survivorship of immature and adult stages, using life-table analysis. We found that in larval habitats located in forested areas, only 4-9% of first-instar larvae developed into adults and the development length exceeded 20 days. Mean water temperature of aquatic habitats in the deforested area was 4.8-6.1 degrees C higher than that in the forested area, larval-to-adult survivorship was increased to 65-82%, and larval-to-adult development time was shortened by 8-9 days. The average indoor temperature in houses in the deforested area was 1.7-1.8 degrees C higher than in the forested area, and the relative humidity was 22-25% lower. The median survival time of adult mosquitoes in the deforested area was 49-55% higher than those in the forested area. The net reproductive rate of female mosquitoes in the deforested area was 1.7- to 2.6-fold higher than that in the forested area. Compared with previously published data on An. gambiae, the net reproductive rate of An. arabiensis was only 0.8-1.3% of Anopheles gambiae in the forested area and 2.3-2.6% in the deforested area. Therefore, the current ambient climate condition is less permissive to An. arabiensis than to An. gambiae in western Kenya highlands. However, environmental changes such as deforestation and global warming may facilitate the establishment of An. arabiensis populations in the highlands.

Feeding and resting behaviour of malaria vector, Anopheles arabiensis with reference to zooprophylaxis.

PubMed

Mahande, Aneth; Mosha, Franklin; Mahande, Johnson; Kweka, Eliningaya

2007-07-30

The most important factor for effective zooprophylaxis in reducing malaria transmission is a predominant population of a strongly zoophilic mosquito, Anopheles arabiensis. The feeding preference behaviour of Anopheline mosquitoes was evaluated in odour-baited entry trap (OBET). Mosquitoes were captured daily using odour-baited entry traps, light traps and hand catch both indoor and in pit traps. Experimental huts were used for release and recapture experiment. The mosquitoes collected were compared in species abundances. Anopheles arabiensis was found to account for over 99% of Anopheles species collected in the study area in Lower Moshi, Northern Tanzania. In experimental release/capture trials conducted at the Mabogini verandah huts, An. arabiensis was found to have higher exophilic tendency (80.7%) compared to Anopheles gambiae (59.7%) and Culex spp. (60.8%). OBET experiments conducted at Mabogini collected a total of 506 An. arabiensis in four different trials involving human, cattle, sheep, goat and pig. Odours from the cattle attracted 90.3% (243) compared to odours from human, which attracted 9.7% (26) with a significant difference at P = 0.005. Odours from sheep, goat and pig attracted 9.7%, 7.2% and 7.3%, respectively. Estimation of HBI in An. arabiensis collected from houses in three lower Moshi villages indicated lower ratios for mosquitoes collected from houses with cattle compared to those without cattles. HBI was also lower in mosquitoes collected outdoors (0.1-0.3) compared to indoor (0.4-0.9). In discussing the results, reference has been made to observation of exophilic, zoophilic and feeding tendencies of An. arabiensis, which are conducive for zooprophylaxis. It is recommended that in areas with a predominant An. arabiensis population, cattle should be placed close to dwelling houses in order to maximize the effects of zooprophylaxis. Protective effects of human from malaria can further be enhanced by keeping cattle in surroundings of residences.

Attractiveness of MM-X Traps Baited with Human or Synthetic Odor to Mosquitoes (Diptera: Culicidae) in The Gambia

PubMed Central

QIU, YU TONG; SMALLEGANGE, RENATE C.; TER BRAAK, CAJO J. F.; SPITZEN, JEROEN; VAN LOON, JOOP J. A.; JAWARA, MUSA; MILLIGAN, PAUL; GALIMARD, AGNES M.; VAN BEEK, TERIS A.; KNOLS, BART G. J.; TAKKEN, WILLEM

2013-01-01

Chemical cues play an important role in the host-seeking behavior of blood-feeding mosquitoes (Diptera: Culicidae). A field study was carried out in The Gambia to investigate the effects of human odor or synthetic odor blends on the attraction of mosquitoes. MM-X traps baited with 16 odor blends to which carbon dioxide (CO2) was added were tested in four sets of experiments. In a second series of experiments, MM-X traps with 14 odor blends without CO2 were tested. A blend of ammonia and l-lactic acid with or without CO2 was used as control odor in series 1 and 2, respectively. Centers for Disease Control and Prevention (CDC) traps were placed in a traditional house and an experimental house to monitor mosquito densities during the experiments. The MM-X traps caught a total number of 196,756 mosquitoes, with the most abundant species belonging to the genera Mansonia (70.6%), Anopheles (17.5%), and Culex (11.5%). The most abundant mosquito species caught by the CDC traps (56,290 in total) belonged to the genera Mansonia (59.4%), Anopheles (16.0% An. gambiae s.l. Giles, and 11.3% An. ziemanni Grünberg), and Culex (11.6%). MM-X traps baited with synthetic blends were in many cases more attractive than MM-X traps baited with human odors. Addition of CO2 to synthetic odors substantially increased the catch of all mosquito species in the MM-X traps. A blend of ammonia + L-lactic acid + CO2 + 3-methylbutanoic acid was the most attractive odor for most mosquito species. The candidate odor blend shows the potential to enhance trap collections so that traps will provide better surveillance and possible control. PMID:18047195

Characterization and expression analysis of gene encoding heme peroxidase HPX15 in major Indian malaria vector Anopheles stephensi (Diptera: Culicidae).

PubMed

Kajla, Mithilesh; Kakani, Parik; Choudhury, Tania Pal; Gupta, Kuldeep; Gupta, Lalita; Kumar, Sanjeev

2016-06-01

The interaction of mosquito immune system with Plasmodium is critical in determining the vector competence. Thus, blocking the crucial mosquito molecules that regulate parasite development might be effective in controlling the disease transmission. In this study, we characterized a full-length AsHPX15 gene from the major Indian malaria vector Anopheles stephensi. This gene is true ortholog of Anopheles gambiae heme peroxidase AgHPX15 (AGAP013327), which modulates midgut immunity and regulates Plasmodium falciparum development. We found that AsHPX15 is highly induced in mosquito developmental stages and blood fed midguts. In addition, this is a lineage-specific gene that has identical features and 65-99% amino acids identity with other HPX15 genes present in eighteen worldwide-distributed anophelines. We discuss that the conserved HPX15 gene might serve as a common target to manipulate mosquito immunity and arresting Plasmodium development inside the vector host. Copyright © 2016 Elsevier B.V. All rights reserved.

Comparison of transmission parameters between Anopheles argyritarsis and Anopheles pseudopunctipennis in two ecologically different localities of Bolivia

PubMed Central

2013-01-01

Background Anopheles (Anopheles) pseudopunctipennis is a recognized malaria vector in the slopes of the Andes of Bolivia. There, other species might be involved in malaria transmission and one candidate could be Anopheles argyritarsis. Although it is generally admitted that this species is not a malaria vector in the neotropical region, its potential role in transmission is still controversial and this situation has to be cleared, at least for Bolivia. Comparing the vectorial efficiency of An. pseudopunctipennis with that of An. argyritarsis could solve the question. Methods The two species were sampled throughout Bolivia to estimate their degree of co-existence in their distribution range. Vectorial efficiencies of the two species were compared in two ecologically different localities where the species were sympatric by analysing their vectorial capacities and components (i e, human biting rates, human biting index, survival, durations of the gonotrophic cycle and extrinsic cycle), and the entomological inoculation rates (EIR). Mosquitoes were sampled monthly during more than one year in the two localities. A monthly sample consisted in hourly captures in four houses (inside and outside) in each locality, during four consecutive nights. Climatic variables (temperature, humidity, potential evapo-transpiration and precipitations) were recorded to better understand variability in the entomological parameters. Relationships were analysed using multivariate methods. Results Anopheles pseudopunctipennis and An. argyritarsis are “altitude” species, sharing the same geographical distribution range in the Andes of Bolivia. No Plasmodium parasite was identified in An. argyritarsis and estimates of the vectorial capacity indicated that it is not a malaria vector in the two studied localities, unlike An. pseudopunctipennis which showed positive EIRs. This latter species, although not a very good malaria vector, exhibited better life traits values and better behavioural

Light manipulation of mosquito behaviour: acute and sustained photic suppression of biting activity in the Anopheles gambiae malaria mosquito.

PubMed

Sheppard, Aaron D; Rund, Samuel S C; George, Gary F; Clark, Erin; Acri, Dominic J; Duffield, Giles E

2017-06-16

Host-seeking behaviours in anopheline mosquitoes are time-of-day specific, with a greater propensity for nocturnal biting. We investigated how a short exposure to light presented during the night or late day can inhibit biting activity and modulate flight activity behaviour. Anopheles gambiae (s.s.), maintained on a 12:12 LD cycle, were exposed transiently to white light for 10-min at the onset of night and the proportion taking a blood meal in a human biting assay was recorded every 2 h over an 8-h duration. The pulse significantly reduced biting propensity in mosquitoes 2 h following administration, in some trials for 4 h, and with no differences detected after 6 h. Conversely, biting levels were significantly elevated when mosquitoes were exposed to a dark treatment during the late day, suggesting that light suppresses biting behaviour even during the late daytime. These data reveal a potent effect of a discrete light pulse on biting behaviour that is both immediate and sustained. We expanded this approach to develop a method to reduce biting propensity throughout the night by exposing mosquitoes to a series of 6- or 10-min pulses presented every 2 h. We reveal both an immediate suppressive effect of light during the exposure period and 2 h after the pulse. This response was found to be effective during most times of the night: however, differential responses that were time-of-day specific suggest an underlying circadian property of the mosquito physiology that results in an altered treatment efficacy. Finally, we examined the immediate and sustained effects of light on mosquito flight activity behaviour following exposure to a 30-min pulse, and observed activity suppression during early night, and elevated activity during the late night. As mosquitoes and malaria parasites are becoming increasingly resistant to insecticide and drug treatment respectively, there is a necessity for the development of innovative control strategies beyond insecticide

Situation Report--Argentina, France, Gambia, Grenada, Hungary, Nepal, and Paraguay.

ERIC Educational Resources Information Center

International Planned Parenthood Federation, London (England).

Data relating to population and family planning in seven foreign countries are presented in these situation reports. Countries included are Argentina, France, Gambia, Grenada, Hungary, Nepal, and Paraguay. Information is provided, where appropriate and available, under two topics, general background and family planning situation. General…

Synergy between Repellents and Organophosphates on Bed Nets: Efficacy and Behavioural Response of Natural Free-Flying An. gambiae Mosquitoes

PubMed Central

Pennetier, Cédric; Costantini, Carlo; Corbel, Vincent; Licciardi, Séverine; Dabiré, Roch K.; Lapied, Bruno; Chandre, Fabrice; Hougard, Jean-Marc

2009-01-01

Background Chemicals are used on bed nets in order to prevent infected bites and to kill aggressive malaria vectors. Because pyrethroid resistance has become widespread in the main malaria vectors, research for alternative active ingredients becomes urgent. Mixing a repellent and a non-pyrethroid insecticide seemed to be a promising tool as mixtures in the laboratory showed the same features as pyrethroids. Methodology/Principal Findings We present here the results of two trials run against free-flying Anopheles gambiae populations comparing the effects of two insect repellents (either DEET or KBR 3023, also known as icaridin) and an organophosphate insecticide at low-doses (pirimiphos-methyl, PM) used alone and in combination on bed nets. We showed that mixtures of PM and the repellents induced higher exophily, blood feeding inhibition and mortality among wild susceptible and resistant malaria vectors than compounds used alone. Nevertheless the synergistic interactions are only involved in the high mortality induced by the two mixtures. Conclusion These field trials argue in favour of the strategy of mixing repellent and organophosphate on bed nets to better control resistant malaria vectors. PMID:19936249

Risk factors for house-entry by malaria vectors in a rural town and satellite villages in The Gambia

PubMed Central

Kirby, Matthew J; Green, Clare; Milligan, Paul M; Sismanidis, Charalambos; Jasseh, Momadou; Conway, David J; Lindsay, Steven W

2008-01-01

Background In the pre-intervention year of a randomized controlled trial investigating the protective effects of house screening against malaria-transmitting vectors, a multi-factorial risk factor analysis study was used to identify factors that influence mosquito house entry. Methods Mosquitoes were sampled using CDC light traps in 976 houses, each on one night, in Farafenni town and surrounding villages during the malaria-transmission season in The Gambia. Catches from individual houses were both (a) left unadjusted and (b) adjusted relative to the number of mosquitoes caught in four sentinel houses that were operated nightly throughout the period, to allow for night-to-night variation. Houses were characterized by location, architecture, human occupancy and their mosquito control activities, and the number and type of domestic animals within the compound. Results 106,536 mosquitoes were caught, of which 55% were Anopheles gambiae sensu lato, the major malaria vectors in the region. There were seven fold higher numbers of An. gambiae s.l. in the villages (geometric mean per trap night = 43.7, 95% confidence intervals, CIs = 39.5–48.4) than in Farafenni town (6.3, 5.7–7.2) and significant variation between residential blocks (p < 0.001). A negative binomial multivariate model performed equally well using unadjusted or adjusted trap data. Using the unadjusted data the presence of nuisance mosquitoes was reduced if the house was located in the town (odds ratio, OR = 0.11, 95% CIs = 0.09–0.13), the eaves were closed (OR = 0.71, 0.60–0.85), a horse was tethered near the house (OR = 0.77, 0.73–0.82), and churai, a local incense, was burned in the room at night (OR = 0.56, 0.47–0.66). Mosquito numbers increased per additional person in the house (OR = 1.04, 1.02–1.06) or trapping room (OR = 1.19, 1.13–1.25) and when the walls were made of mud blocks compared with concrete (OR = 1.44, 1.10–1.87). Conclusion This study demonstrates that the risk of

Impact of insecticide-treated bed nets on malaria transmission indices on the south coast of Kenya

PubMed Central

2011-01-01

Background Besides significantly reducing malaria vector densities, prolonged usage of bed nets has been linked to decline of Anopheles gambiae s.s. relative to Anopheles arabiensis, changes in host feeding preference of malaria vectors, and behavioural shifts to exophagy (outdoor biting) for the two important malaria vectors in Africa, An. gambiae s.l. and Anopheles funestus. In southern coastal Kenya, bed net use was negligible in 1997-1998 when Anopheles funestus and An. gambiae s.s. were the primary malaria vectors, with An. arabiensis and Anopheles merus playing a secondary role. Since 2001, bed net use has increased progressively and reached high levels by 2009-2010 with corresponding decline in malaria transmission. Methods To evaluate the impact of the substantial increase in household bed net use within this area on vector density, vector composition, and human-vector contact, indoor and outdoor resting mosquitoes were collected in the same region during 2009-2010 using pyrethrum spray catches and clay pots for indoor and outdoor collections respectively. Information on bed net use per sleeping spaces and factors influencing mosquito density were determined in the same houses using Poisson regression analysis. Species distribution was determined, and number of mosquitoes per house, human-biting rates (HBR), and entomological inoculation rate (EIR) were compared to those reported for the same area during 1997-1998, when bed net coverage had been minimal. Results Compared to 1997-1998, a significant decline in the relative proportion of An. gambiae s.s. among collected mosquitoes was noted, coupled with a proportionate increase of An. arabiensis. Following > 5 years of 60-86% coverage with bed nets, the density, human biting rate and EIR of indoor resting mosquitoes were reduced by more than 92% for An. funestus and by 75% for An. gambiae s.l. In addition, the host feeding choice of both vectors shifted more toward non-human vertebrates. Besides bed net use

The phylogenetic relationships of known mosquito (Diptera: Culicidae) mitogenomes.

PubMed

Chu, Hongliang; Li, Chunxiao; Guo, Xiaoxia; Zhang, Hengduan; Luo, Peng; Wu, Zhonghua; Wang, Gang; Zhao, Tongyan

2018-01-01

The known mosquito mitogenomes, containing a total of 34 species, which belong to five genera, were collected from GenBank, and the practicality and effectiveness of the variation in the complete mitochondrial DNA genome and portions of mitochondrial COI gene were assessed to reconstruct the phylogeny of mosquitoes. Phylogenetic trees were reconstructed on the basis of parsimony, maximum likelihood, and Bayesian (BI) methods. It is concluded that: (1) Both mitogenomes and COI gene support the monophly of following taxa: Subgenus Nyssorhynchus, Subgenus Cellia, Anopheles albitarsis complex, Anopheles gambiae complex, and Anopheles punctulatus group; (2) Genus Aedes is not monophyletic relative to Ochlerotatus vigilax; (3) The mitogenome results indicate a close relationship between Anopheles epiroticus and Anopheles gambiae complex, Anopheles dirus complex and Anopheles punctulatus group, respectively; (4) The Bayesian posterior probability (BPP) within phylogenetic tree reconstructed by mitogenomes is higher than COI tree. The results show that phylogenetic relationships reconstructed using the mitogenomes were more similar to those based on morphological data.

Anopheles (Anopheles) pseudopunctipennis Theobald (Diptera: Culicidae): Neotype Designation and Description

DTIC Science & Technology

2004-01-01

de- scribed from different areas of South America, includ- ing Ecuador, Colombia, Chile , and Argentina. Three names thatwere applied to this species...Especiales de Salud, Bogota, Colombia (DMB), Instituto Biologie “Juan Noe,” Santiago, Chile (IZC), Museo Argentino de Ciencias Naturales, Buenos Aires...Anopheles) pseudopunctipennis ssp. neghmei Mann 1950: 34 (M, F*, L*, E*). Type: Ouebrada de Minemine, Tarapaca, Chile (IZC). Anopheles (Anopheles

Monitoring pyrethroid insecticide resistance in major malaria vector Anopheles culicifacies: comparison of molecular tools and conventional susceptibility test.

PubMed

Djadid, Navid Dinparast; Forouzesh, Flora; Karimi, Mohsen; Raeisi, Ahmad; Hassan-Zehi, Abdoulghaffar; Zakeri, Sedigheh

2007-07-01

Anopheles culicifacies is a main malaria vector in southeastern part of Iran, bordring Afghanistan and Pakistan. So far, resistance to DDT, dieldrin, malathion and partial tolerance to pyrethroids has been reported in An. stephensi, but nothing confirmed on resistance status of An. culicifacies in Iran. In current study, along with WHO routine susceptibility test with DDT (4%), dieldrin (0.4%), malathion (5%), permethrin (0.25%), lambadacyhalothrin (0.1%), and deltamethrin 0.025, we cloned and sequenced segment VI of domain II (SII6) in voltage-gated sodium channel (vgsc) gene of An. culicifacies specimens collected in Sistan and Baluchistan province (Iran). A 221-bp amplified fragment showed 91% and 93% similarity with exon I and exon II of An. gambiae. The size of intron II in An. culicifacies is 62 bp, while in An. gambiae is 57 bp. The major difference within An. culicifacies specimens and also with An. gambiae is in position 29 of exon I, which led to substitution of Leu to His amino acid. This data will act as first report on partial sequence of vgsc gene and its polymorphism in An. culicifacies. A Leu to His amino acid substitution detected upstream the formerly known knockdown resistance (kdr) mutation site could be an indication for other possible mutations related to insecticide resistance. However, the result of WHO susceptibility test carried out in Baluchistan of Iran revealed a level of tolerance to DDT and dieldrin, but almost complete susceptibility to pyrethroids in An. culicifacies. We postulate that the molecular diagnostic tool developed for detection and identification of kdr-related mutations in An. culicifacies, could be useful in monitoring insecticide resistance in Iran and neighbouring countries such as Pakistan and Afghanistan. A phylogenetic tree also constructed based on the sequence of exon I and II, which readily separated An. culicifacies populations from An. stephensi, An. fluviatilis and An. gambiae.

Potential mosquito repellent compounds of Ocimum species against 3N7H and 3Q8I of Anopheles gambiae.

PubMed

Gaddaguti, Venugopal; Venkateswara Rao, Talluri; Prasada Rao, Allu

2016-06-01

Mosquitoes are exceptionally efficient in detecting their hosts for blood meal using odorant binding proteins, viz. 3N7H and 3Q8I and spread several dreadful diseases. DEET is a synthetic mosquito repellent widely used all over world for protection against mosquito bite. Reports reveal that, synthetic mosquito repellents may pose health problems in considerably large population. In view of the above fact, we made an attempt to discover efficient and novel natural mosquito repellent compounds with least impact on human health. Methanolic leaf extracts of Ocimum basilicum Linn. var. pilosum (willd.)-Benth and Ocimum tenuiflorum var. CIM-AYU were subjected to GC-MS analysis and obtained 35 phytochemical constituents. Repellent potentiality of the Ocimum compounds was assessed against 3Q8I and 3N7H of Anopheles gambiae. PDB structures of mosquito odorant binding proteins were downloaded, processed and docking studies were performed along with reference ligand DEET using Schrodinger MAESTRO 9.2 software. Molecular docking results reveal that phenol, 2-methoxy-3-(2-propenyl)-, licopersin, gamma sitosterol and benzene, 1,2-dimethoxy-4-(2-propenyl)- from O. tenuiflorum var. CIM-AYU are strongly bound with 3N7H. Whereas, 4h-1-benzopyran-4-one, 5-hydroxy-6,7-dimethoxy-2-(4-methoxyphenyl)-, catechol and monoacetin from O. basilicum Linn. var. pilosum (willd.)-Benth. show high binding affinity with odorant binding protein 3Q8I. All natural compounds tested in the present study display better docking scores than DEET. The results further substantiate that the 12 out of 35 compounds of the two Ocimum species found to be ideal candidates for design and development of potential mosquito repellents. ADME properties of the tested compounds further confirm that bioactive compounds of Ocimum species were found to be in acceptable range. Synchronized application of at least two different natural compounds (with best docking scores) which target 3N7H and 3Q8I (Odorant Binding Proteins

Malaria vector composition and insecticide susceptibility status in Guinea Conakry, West Africa.

PubMed

Vezenegho, S B; Brooke, B D; Hunt, R H; Coetzee, M; Koekemoer, L L

2009-12-01

This study provides data on malaria vector species composition and insecticide susceptibility status from three localities in Guinea Conakry. A total of 497 mosquitoes were collected resting indoors and morphologically identified as belonging to the Anopheles gambiae complex. The majority of these were An. gambiae s.s. (99.6%), but a small percentage (0.4%) were identified as Anopheles arabiensis. Thirty-four Anopheles funestus s.s. were also collected. The molecular S form of An. gambiae s.s. was predominant over the M form in Siguiri (95%) and Boffa (97.4%), whereas at Mt Nimba the M form was more abundant (61.4%) than the S form (38.1%). One hybrid M/S specimen was recorded from Mt Nimba. Siguiri populations showed high levels of resistance to DDT, dieldrin and bendiocarb. Anopheles gambiae from Boffa were largely susceptible to the insecticides tested. At Mt Nimba, resistance to DDT and bendicocarb was detected. Biochemical enzyme analysis showed that an altered acetylcholinesterase is operating in the field at low levels. The frequency of the 1014F kdr allele in the An. gambiae S form was 0.24 at Siguiri and 0.14 at Mt Nimba. A single RR specimen was found in the M form. The heterogeneity in species composition and resistance profiles between sites requires vector control interventions to be tailored to each site based on the data collected from ongoing monitoring and surveillance.

An Evolution-Based Screen for Genetic Differentiation between Anopheles Sister Taxa Enriches for Detection of Functional Immune Factors

PubMed Central

Takashima, Eizo; Williams, Marni; Eiglmeier, Karin; Pain, Adrien; Guelbeogo, Wamdaogo M.; Gneme, Awa; Brito-Fravallo, Emma; Holm, Inge; Lavazec, Catherine; Sagnon, N’Fale; Baxter, Richard H.; Riehle, Michelle M.; Vernick, Kenneth D.

2015-01-01

Nucleotide variation patterns across species are shaped by the processes of natural selection, including exposure to environmental pathogens. We examined patterns of genetic variation in two sister species, Anopheles gambiae and Anopheles coluzzii, both efficient natural vectors of human malaria in West Africa. We used the differentiation signature displayed by a known coordinate selective sweep of immune genes APL1 and TEP1 in A. coluzzii to design a population genetic screen trained on the sweep, classified a panel of 26 potential immune genes for concordance with the signature, and functionally tested their immune phenotypes. The screen results were strongly predictive for genes with protective immune phenotypes: genes meeting the screen criteria were significantly more likely to display a functional phenotype against malaria infection than genes not meeting the criteria (p = 0.0005). Thus, an evolution-based screen can efficiently prioritize candidate genes for labor-intensive downstream functional testing, and safely allow the elimination of genes not meeting the screen criteria. The suite of immune genes with characteristics similar to the APL1-TEP1 selective sweep appears to be more widespread in the A. coluzzii genome than previously recognized. The immune gene differentiation may be a consequence of adaptation of A. coluzzii to new pathogens encountered in its niche expansion during the separation from A. gambiae, although the role, if any of natural selection by Plasmodium is unknown. Application of the screen allowed identification of new functional immune factors, and assignment of new functions to known factors. We describe biochemical binding interactions between immune proteins that underlie functional activity for malaria infection, which highlights the interplay between pathogen specificity and the structure of immune complexes. We also find that most malaria-protective immune factors display phenotypes for either human or rodent malaria, with

Hydrological modeling of geophysical parameters of arboviral and protozoan disease vectors in Internally Displaced People camps in Gulu, Uganda

PubMed Central

Jacob, Benjamin G; Muturi, Ephantus J; Caamano, Erick X; Gunter, James T; Mpanga, Enoch; Ayine, Robert; Okelloonen, Joseph; Nyeko, Jack Pen-Mogi; Shililu, Josephat I; Githure, John I; Regens, James L; Novak, Robert J; Kakoma, Ibulaimu

2008-01-01

Background The aim of this study was to determine if remotely sensed data and Digital Elevation Model (DEM) can test relationships between Culex quinquefasciatus and Anopheles gambiae s.l. larval habitats and environmental parameters within Internally Displaced People (IDP) campgrounds in Gulu, Uganda. A total of 65 georeferenced aquatic habitats in various IDP camps were studied to compare the larval abundance of Cx. quinquefasciatus and An. gambiae s.l. The aquatic habitat dataset were overlaid onto Land Use Land Cover (LULC) maps retrieved from Landsat imagery with 150 m × 150 m grid cells stratified by levels of drainage. The LULC change was estimated over a period of 14 years. Poisson regression analyses and Moran's I statistics were used to model relationships between larval abundance and environmental predictors. Individual larval habitat data were further evaluated in terms of their covariations with spatial autocorrelation by regressing them on candidate spatial filter eigenvectors. Multispectral QuickBird imagery classification and DEM-based GIS methods were generated to evaluate stream flow direction and accumulation for identification of immature Cx. quinquefasciatus and An. gambiae s.l. and abundance. Results The main LULC change in urban Gulu IDP camps was non-urban to urban, which included about 71.5 % of the land cover. The regression models indicate that counts of An. gambiae s.l. larvae were associated with shade while Cx. quinquefasciatus were associated with floating vegetation. Moran's I and the General G statistics for mosquito density by species and instars, identified significant clusters of high densities of Anopheles; larvae, however, Culex are not consistently clustered. A stepwise negative binomial regression decomposed the immature An. gambiae s.l. data into empirical orthogonal bases. The data suggest the presence of roughly 11% to 28 % redundant information in the larval count samples. The DEM suggest a positive correlation for Culex

Polymorphism of intron-1 in the voltage-gated sodium channel gene of Anopheles gambiae s.s. populations from Cameroon with emphasis on insecticide knockdown resistance mutations.

PubMed

Etang, Josiane; Vicente, Jose L; Nwane, Philippe; Chouaibou, Mouhamadou; Morlais, Isabelle; Do Rosario, Virgilio E; Simard, Frederic; Awono-Ambene, Parfait; Toto, Jean Claude; Pinto, Joao

2009-07-01

Sequence variation at the intron-1 of the voltage-gated sodium channel gene in Anopheles gambiae M- and S-forms from Cameroon was assessed to explore the number of mutational events originating knockdown resistance (kdr) alleles. Mosquitoes were sampled between December 2005 and June 2006 from three geographical areas: (i) Magba in the western region; (ii) Loum, Tiko, Douala, Kribi, and Campo along the Atlantic coast; and (iii) Bertoua, in the eastern continental plateau. Both 1014S and 1014F kdr alleles were found in the S-form with overall frequencies of 14% and 42% respectively. Only the 1014F allele was found in the M-form at lower frequency (11%). Analysis of a 455 bp region of intron-1 upstream the kdr locus revealed four independent mutation events originating kdr alleles, here named MS1 -1014F, S1-1014S and S2-1014S kdr-intron-1 haplotypes in S-form and MS3-1014F kdr-intron-1 haplotype in the M-form. Furthermore, there was evidence for mutual introgression of kdr 1014F allele between the two molecular forms, MS1 and MS3 being widely shared by them. Although no M/S hybrid was observed in analysed samples, this wide distribution of haplotypes MS1 and MS3 suggests inter-form hybridizing at significant level and emphasizes the rapid diffusion of the kdr alleles in Africa. The mosaic of genetic events found in Cameroon is representative of the situation in the West-Central African region and highlights the importance of evaluating the spatial and temporal evolution of kdr alleles for a better management of insecticide resistance.

Explaining variation in adult Anopheles indoor resting abundance: the relative effects of larval habitat proximity and insecticide-treated bed net use.

PubMed

McCann, Robert S; Messina, Joseph P; MacFarlane, David W; Bayoh, M Nabie; Gimnig, John E; Giorgi, Emanuele; Walker, Edward D

2017-07-17

Spatial determinants of malaria risk within communities are associated with heterogeneity of exposure to vector mosquitoes. The abundance of adult malaria vectors inside people's houses, where most transmission takes place, should be associated with several factors: proximity of houses to larval habitats, structural characteristics of houses, indoor use of vector control tools containing insecticides, and human behavioural and environmental factors in and near houses. While most previous studies have assessed the association of larval habitat proximity in landscapes with relatively low densities of larval habitats, in this study these relationships were analysed in a region of rural, lowland western Kenya with high larval habitat density. 525 houses were sampled for indoor-resting mosquitoes across an 8 by 8 km study area using the pyrethrum spray catch method. A predictive model of larval habitat location in this landscape, previously verified, provided derivations of indices of larval habitat proximity to houses. Using geostatistical regression models, the association of larval habitat proximity, long-lasting insecticidal nets (LLIN) use, house structural characteristics (wall type, roof type), and peridomestic variables (cooking in the house, cattle near the house, number of people sleeping in the house) with mosquito abundance in houses was quantified. Vector abundance was low (mean, 1.1 adult Anopheles per house). Proximity of larval habitats was a strong predictor of Anopheles abundance. Houses without an LLIN had more female Anopheles gambiae s.s., Anopheles arabiensis and Anopheles funestus than houses where some people used an LLIN (rate ratios, 95% CI 0.87, 0.85-0.89; 0.84, 0.82-0.86; 0.38, 0.37-0.40) and houses where everyone used an LLIN (RR, 95% CI 0.49, 0.48-0.50; 0.39, 0.39-0.40; 0.60, 0.58-0.61). Cooking in the house also reduced Anopheles abundance across all species. The number of people sleeping in the house, presence of cattle near the house

Détermination des paramètres bioécologiques et entomologiques d’Anopheles gambiae sl dans la transmission du paludisme à Bandundu-ville, République Démocratique de Congo

PubMed Central

Matubi, Emery Metelo; Bukaka, Eric; Luemba, Trésor Bakambana; Situakibanza, Hyppolite; Sangaré, Ibrahim; Mesia, Gauthier; Ngoyi, Dieudonné Mumba; Maniania, Nguya Kalemba; Akikwa, Charles Ngandote; Kanza, Jean Pierre Basilua; Tamfum, Jean-Jacques Muyembe; sudi, Jonas Nagahuedi Bongo

2015-01-01

Introduction La présente étude a été menée à Bandundu-ville (RDC) en vue d'identifier les paramètres écologiques et entomologiques modulant la transmission du paludisme ainsi que leur tendance saisonnière dans cette agglomération. Méthodes Cette étude a été réalisée dans la période du 1er juin au 31 décembre 2011. Des prospections des gîtes larvaires d'anophèles avec récolte ont été réalisées, les paramètres physiques, physico-chimiques et environnementaux déterminés. La densité larvaire a été estimée selon une échelle de classes de densité, inspirée de la méthode de Carron pour chaque type de gîtes. Quarante-huit maisons ont été sélectionnées et prospectées pour la récolte des moustiques par pulvérisation intradomicilaire. L'identification des moustiques a été faite sur base des critères morphologiques de Gilles et Demeillon. L'Indice sporozoïtique (Is) a été déterminé par le test ELISA CSP de Plasmodium falciparum à l'Institut National de Recherche Biomédicale selon le protocole de Robert Wirtz. Les autres paramètres entomologiques comme la densité, le taux d'agressivité, le taux d'inoculation entomologique (TIE) ainsi que l'indice de stabilité ont été déterminés selon le protocole de l'OMS. La régression linéaire a été réalisée au seuil de signification de 0,05 pour identifier les déterminants de la densité larvaire. Résultats Cent-sept gîtes larvaires ont été identifiés et caractérisés en 5 types (digues et puits d'eau, collections d'eau maraîchère et concasseurs moellons, marais Régie de distribution d'eau, marais le long des rivières et ruisseaux et flaques d'eau de pluies). La densité larvaire moyenne a été de 117,4±64,1. Quatre mille cinq cents quatre-vingt-huit moustiques ont été capturés et identifiés, parmi lesquels 1.258 Anopheles gambiae sl avec une densité de 8,86, un taux d'agressivité de 1,55 piqûre par homme par nuit, l'Is de 5,6%, un TIE de 0,085 piq

Anopheles gambiae Circumsporozoite Protein–Binding Protein Facilitates Plasmodium Infection of Mosquito Salivary Glands

PubMed Central

Wang, Jiuling; Zhang, Yue; Zhao, Yang O.; Li, Michelle W. M.; Zhang, Lili; Dragovic, Srdjan; Abraham, Nabil M.; Fikrig, Erol

2013-01-01

Malaria, a mosquito-borne disease caused by Plasmodium species, causes substantial morbidity and mortality throughout the world. Plasmodium sporozoites mature in oocysts formed in the mosquito gut wall and then invade the salivary glands, where they remain until transmitted to the vertebrate host during a mosquito bite. The Plasmodium circumsporozoite protein (CSP) binds to salivary glands and plays a role in the invasion of this organ by sporozoites. We identified an Anopheles salivary gland protein, named CSP-binding protein (CSPBP), that interacts with CSP. Downregulation of CSPBP in mosquito salivary glands inhibited invasion by Plasmodium organisms. In vivo bioassays showed that mosquitoes that were fed blood with CSPBP antibody displayed a 25% and 90% reduction in the parasite load in infected salivary glands 14 and 18 days after the blood meal, respectively. These results suggest that CSPBP is important for the infection of the mosquito salivary gland by Plasmodium organisms and that blocking CSPBP can interfere with the Plasmodium life cycle. PMID:23801601

The impact of livestock on the abundance, resting behaviour and sporozoite rate of malaria vectors in southern Tanzania.

PubMed

Mayagaya, Valeriana S; Nkwengulila, Gamba; Lyimo, Issa N; Kihonda, Japheti; Mtambala, Hassan; Ngonyani, Hassan; Russell, Tanya L; Ferguson, Heather M

2015-01-21

Increases in the coverage of long-lasting insecticidal nets (LLINs) have significantly reduced the abundance of Anopheles gambiae sensu stricto in several African settings, leaving its more zoophagic sibling species Anopheles arabiensis as the primary vector. This study investigated the impact of livestock ownership at the household level on the ecology and malaria infection rate of vectors in an area of Tanzania where An. arabiensis accounts for most malaria transmission. Mosquito vectors were collected resting inside houses, animal sheds and in outdoor resting boxes at households with and without livestock over three years in ten villages of the Kilombero Valley, Tanzania. Additionally, the abundance and sporozoite rate of vectors attempting to bite indoors at these households was assessed as an index of malaria exposure. The mean abundance of An. gambiae s.l. biting indoors was similar at houses with and without livestock. In all years but one, the relative proportion of An. arabiensis within the An. gambiae s.l. species complex was higher at households with livestock. Livestock presence had a significant impact on malaria vector feeding and resting behaviour. Anopheles arabiensis were generally found resting in cattle sheds where livestock were present, and inside houses when absent. Correspondingly, the human blood index of An. arabiensis and An. funestus s.l. was significant reduced at households with livestock, whereas that of An. gambiae s.s. was unaffected. Whilst there was some evidence that sporozoite rates within the indoor-biting An. gambiae s.l population was significantly reduced at households with livestock, the significance of this effect varied depending on how background spatial variation was accounted for. These results confirm that the presence of cattle at the household level can significantly alter the local species composition, feeding and resting behaviour of malaria vectors. However, the net impact of this livestock-associated variation in

Optimized Pan-species and Speciation Duplex Real-time PCR Assays for Plasmodium Parasites Detection in Malaria Vectors

PubMed Central

Sandeu, Maurice Marcel; Moussiliou, Azizath; Moiroux, Nicolas; Padonou, Gilles G.; Massougbodji, Achille; Corbel, Vincent; Tuikue Ndam, Nicaise

2012-01-01

Background An accurate method for detecting malaria parasites in the mosquito’s vector remains an essential component in the vector control. The Enzyme linked immunosorbent assay specific for circumsporozoite protein (ELISA-CSP) is the gold standard method for the detection of malaria parasites in the vector even if it presents some limitations. Here, we optimized multiplex real-time PCR assays to accurately detect minor populations in mixed infection with multiple Plasmodium species in the African malaria vectors Anopheles gambiae and Anopheles funestus. Methods Complementary TaqMan-based real-time PCR assays that detect Plasmodium species using specific primers and probes were first evaluated on artificial mixtures of different targets inserted in plasmid constructs. The assays were further validated in comparison with the ELISA-CSP on 200 field caught Anopheles gambiae and Anopheles funestus mosquitoes collected in two localities in southern Benin. Results The validation of the duplex real-time PCR assays on the plasmid mixtures demonstrated robust specificity and sensitivity for detecting distinct targets. Using a panel of mosquito specimen, the real-time PCR showed a relatively high sensitivity (88.6%) and specificity (98%), compared to ELISA-CSP as the referent standard. The agreement between both methods was “excellent” (κ = 0.8, P<0.05). The relative quantification of Plasmodium DNA between the two Anopheles species analyzed showed no significant difference (P = 0, 2). All infected mosquito samples contained Plasmodium falciparum DNA and mixed infections with P. malariae and/or P. ovale were observed in 18.6% and 13.6% of An. gambiae and An. funestus respectively. Plasmodium vivax was found in none of the mosquito samples analyzed. Conclusion This study presents an optimized method for detecting the four Plasmodium species in the African malaria vectors. The study highlights substantial discordance with traditional ELISA-CSP pointing out the

Ecology of urban malaria vectors in Niamey, Republic of Niger.

PubMed

Labbo, Rabiou; Fandeur, Thierry; Jeanne, Isabelle; Czeher, Cyril; Williams, Earle; Arzika, Ibrahim; Soumana, Amadou; Lazoumar, Ramatoulaye; Duchemin, Jean-Bernard

2016-06-08

Urbanization in African cities has major impact on malaria risk. Niamey, the capital of the Republic of Niger, is situated in the West African Sahel zone. The short rainy season and human activities linked with the Niger River influence mosquito abundance. This study aimed at deciphering the factors of distribution of urban malaria vectors in Niamey. The distribution of mosquito aquatic stages was investigated monthly from December 2002 to November 2003, at up to 84 breeding sites, throughout Niamey. An exploratory analysis of association between mosquito abundance and environmental factors was performed by a Principal Component Analysis and confirmed by Kruskall-Wallis non-parametric test. To assess the relative importance of significant factors, models were built for Anopheles and Culicinae. In a second capture session, adult mosquitoes were collected weekly with pyrethrum sprays and CDC light-traps from June 2008 to June 2009 in two differentiated urban areas chosen after the study's first step. Members of the Anopheles gambiae complex were genotyped and Anopheles females were tested for the presence of Plasmodium falciparum circumsporozoite antigens using ELISA. In 2003, 29 % of 8420 mosquitoes collected as aquatic stages were Anopheles. They were significantly more likely to be found upstream, relatively close to the river and highly productive in ponds. These factors remained significant in regression and generalized linear models. The Culicinae were found significantly more likely close to the river, and in the main temporary affluent stream. In 2009, Anopheles specimens, including Anopheles gambiae s.l. (95 %), but also Anopheles funestus (0.6 %) accounted for 18 % of the adult mosquito fauna, with a large difference between the two sampled zones. Three members of the An. gambiae complex were found: Anopheles arabiensis, Anopheles coluzzii, and An. gambiae. Nineteen (1.3 %) out of 1467 females tested for P. falciparum antigen were found positive. The

Efficacy of long-lasting insecticidal nets in use in Macha, Zambia, against the local Anopheles arabiensis population.

PubMed

Norris, Laura C; Norris, Douglas E

2011-08-31

The mosquito Anopheles arabiensis is the primary vector of Plasmodium falciparum in Macha, Zambia. A major portion of Zambia's current malaria control programme relies on long-lasting insecticide-treated nets (LLINs) and indoor residual spraying (IRS) with insecticides. Currently, the efficacy of these measures against An. arabiensis in Macha is unknown, and previous data has shown that An. arabiensis has continued to feed on human hosts, despite high ITN coverage. It is possible that this could be due to either decreased efficacy of ITNs in used in Macha, or pyrethroid resistance in the vector. F1 offspring of field-collected adult An. arabiensis were tested for insecticide resistance, using CDC bottle bioassays and deltamethrin ITN susceptibility assays. The mosquitoes were characterized for the knock-down resistance (kdr) allele by PCR. LLINs that had been in use for two years in nearby villages were collected and tested for residual deltamethrin concentration and net quality, and were used in bioassays against susceptible colonized Anopheles gambiae s.s. Keele. Additionally, a survey on ITN use and care was conducted among LLIN owners. In the F1 An. arabiensis field population, low levels of resistance to DDT and deltamethrin-treated net material were detected by bioassay, although the knock-down resistance (kdr) allele not present in the population. ITN evaluations revealed high variability in residual deltamethrin concentration, quality of the nets, and mosquito mortality in bioassays. Mortality against An. gambiae s.s. in bioassays was correlated with residual deltamethrin concentration, which was dependent upon the number of washes each net had received. Proper LLIN care was a strong determinant of LLIN efficacy, indicating that education on the importance of LLIN use and care is key when distributing nets. As there is little insecticide resistance in the local vector population, degradation of LLINs most likely allowed for continued human feeding by An

To assess whether indoor residual spraying can provide additional protection against clinical malaria over current best practice of long-lasting insecticidal mosquito nets in The Gambia: study protocol for a two-armed cluster-randomised trial

PubMed Central

2011-01-01

Background Recently, there has been mounting interest in scaling-up vector control against malaria in Africa. It needs to be determined if indoor residual spraying (IRS with DDT) will provide significant marginal protection against malaria over current best practice of long-lasting insecticidal nets (LLINs) and prompt treatment in a controlled trial, given that DDT is currently the most persistent insecticide for IRS. Methods A 2 armed cluster-randomised controlled trial will be conducted to assess whether DDT IRS and LLINs combined provide better protection against clinical malaria in children than LLINs alone in rural Gambia. Each cluster will be a village, or a group of small adjacent villages; all clusters will receive LLINs and half will receive IRS in addition. Study children, aged 6 months to 13 years, will be enrolled from all clusters and followed for clinical malaria using passive case detection to estimate malaria incidence for 2 malaria transmission seasons in 2010 and 2011. This will be the primary endpoint. Exposure to malaria parasites will be assessed using light and exit traps followed by detection of Anopheles gambiae species and sporozoite infection. Study children will be surveyed at the end of each transmission season to estimate the prevalence of Plasmodium falciparum infection and the prevalence of anaemia. Discussion Practical issues concerning intervention implementation, as well as the potential benefits and risks of the study, are discussed. Trial Registration ISRCTN01738840 - Spraying And Nets Towards malaria Elimination (SANTE) PMID:21663656

Examining the impact of larval source management and insecticide-treated nets using a spatial agent-based model of Anopheles gambiae and a landscape generator tool

PubMed Central

2013-01-01

Background Agent-based models (ABMs) have been used to estimate the effects of malaria-control interventions. Early studies have shown the efficacy of larval source management (LSM) and insecticide-treated nets (ITNs) as vector-control interventions, applied both in isolation and in combination. However, the robustness of results can be affected by several important modelling assumptions, including the type of boundary used for landscapes, and the number of replicated simulation runs reported in results. Selection of the ITN coverage definition may also affect the predictive findings. Hence, by replication, independent verification of prior findings of published models bears special importance. Methods A spatially-explicit entomological ABM of Anopheles gambiae is used to simulate the resource-seeking process of mosquitoes in grid-based landscapes. To explore LSM and replicate results of an earlier LSM study, the original landscapes and scenarios are replicated by using a landscape generator tool, and 1,800 replicated simulations are run using absorbing and non-absorbing boundaries. To explore ITNs and evaluate the relative impacts of the different ITN coverage schemes, the settings of an earlier ITN study are replicated, the coverage schemes are defined and simulated, and 9,000 replicated simulations for three ITN parameters (coverage, repellence and mortality) are run. To evaluate LSM and ITNs in combination, landscapes with varying densities of houses and human populations are generated, and 12,000 simulations are run. Results General agreement with an earlier LSM study is observed when an absorbing boundary is used. However, using a non-absorbing boundary produces significantly different results, which may be attributed to the unrealistic killing effect of an absorbing boundary. Abundance cannot be completely suppressed by removing aquatic habitats within 300 m of houses. Also, with density-dependent oviposition, removal of insufficient number of aquatic

Video studies of passage by Anopheles gambiae mosquitoes through holes in a simulated bed net: effects of hole size, hole orientation and net environment.

PubMed

Sutcliffe, James; Colborn, Kathryn L

2015-05-13

Holes in netting provide potential routes for mosquitoes to enter ITNs. Despite this, there is little information on how mosquitoes respond to holes in bed nets and how their responses are affected by hole size, shape and orientation or by ambient conditions around the net. Female Anopheles gambiae (G3) were recorded in a simulated bed net consisting of two sizes of untreated netting-covered behavioural arenas placed above and beside (to simulate the bed net roof and sides respectively) the experimenter who was a source of host cues from 'inside' the net. A round hole of 9 mm or 13 mm diameter was cut into the centre of the netting of each arena. Videos of unfed female mosquitoes in arenas were analysed for time spent flying, walking and standing still and for exit through the hole. The effects of the experimenter on temperature and relative humidity around the simulated net were also measured. Mosquitoes were significantly more active in overhead arenas than in arenas to the side. Hole passage was significantly more likely in smaller arenas than larger ones and for larger holes than smaller ones. In arenas to the side, hole passage rate through small holes was about 50% less likely than what could be explained by area alone. Passage rate through holes in overhead arenas was consistent with hole area. Temperature in arenas did not strongly reflect the experimenter's presence in the simulated net. Relative humidity and absolute humidity in overhead arenas, but not in arenas to the side, were immediately affected by experimenter presence. Higher levels of activity in overhead arenas than in arenas to the side were likely due to the rising heat and humidity plume from the experimenter. Lower than expected passage rates through smaller vertically oriented holes may have been be due to an edge effect that does not apply to horizontally oriented holes. Results suggest that current methods of assessing the importance of physical damage to ITNs may not accurately reflect

[New insight into Anopheles coluzzii Coetzee & Wilkerson, 2013 larval ecology in the Wouri estuary, Littoral-Cameroon].

PubMed

Mbida Mbida, A; Etang, J; Akono Ntonga, P; Eboumbou Moukoko, C; Awono-Ambene, P; Tagne, D; Talipouo, A; Ekoko, W; Binyang, J; Tchoffo, R; Lehman, G; Mimpfoundi, R

2017-05-01

Malaria vectors control is essentially based on the use of insecticides against adult mosquitoes. However because of the development of resistance to insecticides, there is now a renewed interest in the management of larval sources. The aim of the present study was to map and characterize the breeding sites of Anopheles coluzzii in the Wouri river estuary in Cameroon. Larval surveys were carried out between December 2013 and August 2014 in rural areas on the island of Manoka and urban area in Youpwe at Douala. Culicidae breeding sites identified were georeferenced and mapped. Their larval productivity was evaluated by the method of "dipping" and their physicochemical parameters measured by spectrophotometry and oximetry. Culicidae collected larvae were reared in the insectarium to the adult stage. Adult mosquitoes were subjected to morphological identification and those belonging to the Anopheles gambiae complex have subsequently been subjected to molecular identification by the PCR-RFLP technique. A total of 240 breeding sites were geo-referenced in the two sites, including 10 types. Abandoned containers and pools were the most frequent breeding sites respectively in Manoka and in Youpwe. After morphological and molecular identification, eleven mosquito species have been identified. Anopheles coluzzii and Culex quinquefasciatus were the most frequent species respectively in Manoka and in Youpwe. Mosquito density was higher in managed gutters and canoes respectively in Manoka and in Youpwe. Culex and Aedes genus were more frequent in the hollow palm and water wells respectively in Manoka and Youpwe. The productivity of breeding sites varied according to the physicochemical parameters. Species richness varied according to the type of breeding site. Anopheles coluzzii was observed for the first time in Cameroon in water storage containers, tires, discarded containers and canoes. This study highlighted diversity in the type of breeding site of An. coluzzii in the

The sterilizing effect of pyriproxyfen on the malaria vector Anopheles gambiae: physiological impact on ovaries development.

PubMed

Koama, Bayili; Namountougou, Moussa; Sanou, Roger; Ndo, Sévérin; Ouattara, Abdoulaye; Dabiré, Roch K; Malone, David; Diabaté, Abdoulaye

2015-03-04

Adult females An. gambiae were exposed in 3 min cone test to treated nets with PPF before or after they were blood fed. The effects of PPF on ovaries development, females oviposition and eggs hatching were assessed. Both unfed and fed mosquitoes exposed to PPF exhibited nearly complete inhibition of fecundity (70-100%) and fertility (90-100%). After females have been exposed once to PPF, the sterilizing effect on their fecundity was observed over 3 consecutive blood meals suggesting that PPF might have an irreversible sterilizing effect. Observation of the ovaries of exposed females to PPF under microscope revealed that the ovaries failed to develop even after several blood meals. The combination of PPF to pyrethroids on bednets could provide better malaria control tool and prevent the further development and spread of pyrethroid resistance in malaria vectors.

Male accessory gland proteins induce female monogamy in anopheline mosquitoes.

PubMed

Shutt, B; Stables, L; Aboagye-Antwi, F; Moran, J; Tripet, F

2010-03-01

The role of male accessory gland (MAG) secretions in inducing refractoriness to further mating in mosquitoes (Diptera: Culicidae) was established in the late 1960s. In a set of simple experiments, MAG extract was injected intra-thoraxically into the hemocoel of virgin Aedes aegypti (L.), Culex pipiens pipiens (L.) and Anopheles quadrimaculatus Say females. This subsequently caused most females to remain unmated when exposed to males. For anophelines these findings were later challenged by a study involving intra-abdominal injections of MAG extracts into Anopheles gambiae Giles s.l. and Anopheles albimanus Wiedmann females, which failed to induce refractoriness to further mating. These findings led to controversy about the respective role of sperm and accessory gland peptides in inducing female monogamy in Anopheles and are at odds with our current understanding of the mating process in Drosophila spp. (Diptera: Drosophillidae) and other dipterans. Here we confirm the function of MAG secretions in anophelines experimentally by showing that intra-thoracic injections in Anopheles stephensi Liston and in the M and S molecular forms of An. gambiae s.s. result in the expected female monogamy. Cross-injections of MAG extracts between the M and S molecular forms of An. gambiae, two cryptic taxa within An. gambiae s.s. which are thought to be undergoing incipient speciation, also elicited effective refractoriness, suggesting that the two sub-taxa have not diverged with regard to sex peptides responsible for female monogamy. Importantly, this also suggests that the rare cases of re-mating following cross-mating observed in this species may not be a form of reproductive barrier between molecular forms.

The use of sequential mark-release-recapture experiments to estimate population size, survival and dispersal of male mosquitoes of the  Anopheles gambiae complex in Bana, a west African humid savannah village.

PubMed

Epopa, Patric Stephane; Millogo, Abdoul Azize; Collins, Catherine Matilda; North, Ace; Tripet, Frederic; Benedict, Mark Quentin; Diabate, Abdoulaye

2017-08-07

Vector control is a major component of the malaria control strategy. The increasing spread of insecticide resistance has encouraged the development of new tools such as genetic control which use releases of modified male mosquitoes. The use of male mosquitoes as part of a control strategy requires an improved understanding of male mosquito biology, including the factors influencing their survival and dispersal, as well as the ability to accurately estimate the size of a target mosquito population. This study was designed to determine the seasonal variation in population size via repeated mark-release-recapture experiments and to estimate the survival and dispersal of male mosquitoes of the Anopheles gambiae complex in a small west African village. Mark-release-recapture experiments were carried out in Bana Village over two consecutive years, during the wet and the dry seasons. For each experiment, around 5000 (3407-5273) adult male Anopheles coluzzii mosquitoes were marked using three different colour dye powders (red, blue and green) and released in three different locations in the village (centre, edge and outside). Mosquitoes were recaptured at sites spread over the village for seven consecutive days following the releases. Three different capture methods were used: clay pots, pyrethroid spray catches and swarm sampling. Swarm sampling was the most productive method for recapturing male mosquitoes in the field. Population size and survival were estimated by Bayesian analyses of the Fisher-Ford model, revealing an about 10-fold increase in population size estimates between the end of dry season (10,000-50,000) to the wet season (100,000-500,000). There were no detectable seasonal effects on mosquito survival, suggesting that factors other than weather may play an important role. Mosquito dispersal ranged from 40 to 549 m over the seven days of each study and was not influenced by the season, but mainly by the release location, which explained more than 44% of

Insecticide susceptibility and vector status of natural populations of Anopheles arabiensis from Sudan.

PubMed

Abdalla, H; Matambo, T S; Koekemoer, L L; Mnzava, A P; Hunt, R H; Coetzee, M

2008-03-01

Species composition, blood meal source, sporozoite infection rate, insecticide resistance and the kdr mutations were investigated in the Anopheles gambiae complex from 13 sentinel sites in central Sudan. Species identification revealed that 89.5% of 960 specimens were A. arabiensis. Of 310 indoor resting females, 88.1% were found to have fed on humans, while 10.6% had fed on bovines. The overall sporozoite infection rate from the five localities tested was 2.3%, ranging from 0 to 5.5%. Insecticide susceptibility bioassay results showed 100% mortality on bendiocarb, 54.6-94.2% on permethrin, 55.4-99.1% on DDT and 76.8-100% on malathion. The kdr analysis by PCR and sequencing revealed the presence of the Leu-Phe mutation in both permethrin and DDT bioassays. There was no significant difference in the frequency of kdr (P>0.05) between dead and surviving specimens. These findings have serious implications for the malaria control programmes in Gezira and Sennar states.

Mapping insecticide resistance and characterization of resistance mechanisms in Anopheles arabiensis (Diptera: Culicidae) in Ethiopia.

PubMed

Alemayehu, Eba; Asale, Abebe; Eba, Kasahun; Getahun, Kefelegn; Tushune, Kora; Bryon, Astrid; Morou, Evangelia; Vontas, John; Van Leeuwen, Thomas; Duchateau, Luc; Yewhalaw, Delenasaw

2017-09-02

The emergence and spread of insecticide resistance in the major African malaria vectors Anopheles gambiae (s.s.) and An. arabiensis may compromise the current vector control interventions and threatens the global malaria control and elimination efforts. Insecticide resistance was monitored in several study sites in Ethiopia from 2013 to 2015 using papers impregnated with discriminating concentrations of DDT, deltamethrin, bendiocarb, propoxur, malathion, fenitrothion and pirimiphos-methyl, following the WHO insecticide susceptibility test procedure. Mosquitoes sampled from different localities for WHO bioassay were morphologically identified as An. gambiae (s.l.) using standard taxonomic keys. Samples were identified to species using species-specific polymerase chain reaction (PCR) and screened for the presence of target site mutations L1014F, L1014S and N1575Y in the voltage gated sodium channel (VGSC) gene and G119S in the acethylcholinesterase (AChE) gene using allele-specific PCR. Biochemical assays were performed to assess elevated levels of acetylcholinesterases, carboxylcholinesterases, glutathione-S-transferases (GSTs) and cytochrome P450s monooxygenases in wild populations of An. arabiensis, compared to the fully susceptible Sekoru An. arabiensis laboratory strain. Populations of An. arabiensis were resistant to DDT and deltamethrin but were susceptible to fenitrothion in all the study sites. Reduced susceptibility to malathion, pirimiphos-methyl, propoxur and bendiocarb was observed in some of the study sites. Knockdown resistance (kdr L1014F) was detected in all mosquito populations with allele frequency ranging from 42 to 91%. Elevated levels of glutathione-S-transferases (GSTs) were detected in some of the mosquito populations. However, no elevated levels of monooxygenases and esterases were detected in any of the populations assessed. Anopheles arabiensis populations from all surveyed sites in Ethiopia exhibited resistance against DDT and pyrethroids

First Record of Anopheles oryzalimnetes, Anopheles argyritarsis, and Anopheles sawyeri (Diptera: Culicidae) in the Caatinga Biome, Semiarid Scrubland of Sergipe State, Brazil.

PubMed

Marteis, L S; Sallum, M A M; Natal, D; Oliveira, T M P; Gama, R A; Dolabella, S S; Santos, R L C

2015-09-01

Caatinga is one of the least known biomes of Brazil in relation to biodiversity. The dry condition of semiarid areas has been associated in the past with low richness of fauna and flora, not encouraging studies in this region. There is a lack of mosquito records including anophelines. Thus, to investigate the biodiversity of Anopheles mosquitoes in the Caatinga biome, we collected immature mosquitoes in aquatic habitats in a conservation reserve located in the northwestern portion of Sergipe state. The captured specimens were initially identified as Anopheles albitarsis l.s. and Anopheles argyritarsis l.s. To confirm the morphological identification, sequences were generated by cytochrome oxidase subunit I mitocondrial gene. The results showed that the specimens belong to the species Anopheles oryzalimnetes, An. argyritarsis, and Anopheles sawyeri. These are the first records of these species in this region. The presence of Anopheles in the Caatinga biome, which is characterized by arid and semiarid climatic conditions, encourages the interest in the study of biological, physiological, and behavioral adaptations, selected over time, which allow these mosquito populations to survive through the long periods of drought that is characteristic of this region. © The Authors 2015. Published by Oxford University Press on behalf of Entomological Society of America. All rights reserved. For Permissions, please email: journals.permissions@oup.com.

The knowledge, attitudes and practices of wintersun vacationers to the Gambia toward prevention of malaria: is it really that bad?

PubMed Central

2014-01-01

Background Each year clusters of imported malaria cases are observed in Dutch wintersun vacationers returning from The Gambia. To gain more insight in the travel health preparation and awareness of these travellers, the knowledge, attitudes and practices (KAP) of this travel group was studied by analysing the data of the Continuous Dutch Schiphol Airport Survey. Methods In the years 2002 to 2009 a questionnaire-based survey was conducted at the Dutch Schiphol Airport with the aim to study the KAP, i.e. accuracy of risk perception (“knowledge”), intended risk-avoiding behaviour (“attitude”) and use of personal protective measures and malaria chemoprophylaxis (“practice”) toward prevention malaria in travellers to The Gambia. Travellers to other high-risk destinations served as controls. Results The KAP of travellers to The Gambia toward prevention of malaria was significantly better than that observed in other travellers. Trend analyses indicated that attitude improved over time in both groups but knowledge did not change. Only in travellers to high-risk countries other than The Gambia significant increases in protection rates were observed over time. Conclusions The KAP of travellers to The Gambia toward prevention of malaria was better than that observed in travellers to destinations other than The Gambia. Trend analyses revealed a significant improvement of intended risk avoiding behaviour but not in protection rates or risk perception. PMID:24581328

Low prevalence of Plasmodium and absence of malaria transmission in Conakry, Guinea: prospects for elimination.

PubMed

Kouassi, Bernard L; de Souza, Dziedzom K; Goepogui, Andre; Balde, Siradiou M; Diakité, Lamia; Sagno, Arsène; Djameh, Georgina I; Chammartin, Frédérique; Vounatsou, Penelope; Bockarie, Moses J; Utzinger, Jürg; Koudou, Benjamin G

2016-03-18

Over the past 15 years, mortality and morbidity due to malaria have been reduced substantially in sub-Saharan Africa and local elimination has been achieved in some settings. This study addresses the bio-ecology of larval and adult stages of malaria vectors, Plasmodium infection in Anopheles gambiae s.l. in the city of Conakry, Guinea, and discusses the prospect for malaria elimination. Water bodies were prospected to identify potential mosquito breeding sites for 6 days each in the dry season (January 2013) and in the rainy season (August 2013), using the dipping method. Adult mosquitoes were collected in 15 communities in the five districts of Conakry using exit traps and indoor spraying catches over a 1-year period (November 2012 to October 2013). Molecular approaches were employed for identification of Anopheles species, including An. coluzzii and An. gambiae s.s. Individual An. gambiae mosquitoes were tested for Plasmodium falciparum and P. vivax sporozoites using the VecTest™ malaria panel assay and an enzyme-linked immunosorbent assay. A systematic research of Ministry of Health statistical yearbooks was performed to determine malaria prevalence in children below the age of 5 years. Culex larval breeding sites were observed in large numbers throughout Conakry in both seasons. While Anopheles larval breeding sites were less frequent than Culex breeding sites, there was a high odds of finding An. gambiae mosquito larvae in agricultural sites during the rainy season. Over the 1-year study period, a total of 14,334 adult mosquitoes were collected; 14,135 Culex (98.6%) and 161 (1.1%) from the An. gambiae complex. One-hundred and twelve Anopheles mosquitoes, mainly collected from rice fields and gardens, were subjected to molecular analysis. Most of the mosquitoes were An. gambiae s.s. (n = 102; 91.1%) while the remaining 10 (8.9%) were An. melas. The molecular M form of An. gambiae s.s. was predominant (n = 89; 79.5%). The proportions of kdr genotype in the An

Mosquitocidal Activity and Mode of Action of the Isoxazoline Fluralaner

USDA-ARS?s Scientific Manuscript database

Mosquitoes, such as Aedes aegypti and Anopheles gambiae, are important vectors of human diseases. Fluralaner, a recently introduced parasiticide, was evaluated as a mosquitocide in this study.On Ae. aegypti and An. gambiae fourth-instar larvae, fluralaner had 24-h LC50 (lethal concentration for 50% ...

Mosquitocidal activity and mode of action of the isoxazoline fluralaner

USDA-ARS?s Scientific Manuscript database

Mosquitoes, such as Aedes aegypti and Anopheles gambiae, are important vectors of human diseases. Fluralaner, a recently introduced parasiticide, was evaluated as a mosquitocide in this study.On Ae. aegypti and An. gambiae fourth-instar larvae, fluralaner had 24-h LC50 (lethal concentration for 50% ...

Human antibody responses to the Anopheles salivary gSG6-P1 peptide: a novel tool for evaluating the efficacy of ITNs in malaria vector control.

PubMed

Drame, Papa Makhtar; Poinsignon, Anne; Besnard, Patrick; Cornelie, Sylvie; Le Mire, Jacques; Toto, Jean-Claude; Foumane, Vincent; Dos-Santos, Maria Adelaide; Sembène, Mbacké; Fortes, Filomeno; Simondon, Francois; Carnevale, Pierre; Remoue, Franck

2010-12-14

To optimize malaria control, WHO has prioritised the need for new indicators to evaluate the efficacy of malaria vector control strategies. The gSG6-P1 peptide from gSG6 protein of Anopheles gambiae salivary glands was previously designed as a specific salivary sequence of malaria vector species. It was shown that the quantification of human antibody (Ab) responses to Anopheles salivary proteins in general and especially to the gSG6-P1 peptide was a pertinent biomarker of human exposure to Anopheles. The present objective was to validate this indicator in the evaluation of the efficacy of Insecticide Treated Nets (ITNs). A longitudinal evaluation, including parasitological, entomological and immunological assessments, was conducted on children and adults from a malaria-endemic area before and after the introduction of ITNs. Significant decrease of anti-gSG6-P1 IgG response was observed just after the efficient ITNs use. Interestingly, specific IgG Ab level was especially pertinent to evaluate a short-time period of ITNs efficacy and at individual level. However, specific IgG rose back up within four months as correct ITN use waned. IgG responses to one salivary peptide could constitute a reliable biomarker for the evaluation of ITN efficacy, at short- and long-term use, and provide a valuable tool in malaria vector control based on a real measurement of human-vector contact.

Existence of the rdl mutant alleles among the anopheles malaria vector in Indonesia

PubMed Central

2012-01-01

Background The gamma-aminobutyric acid (GABA) receptor-chloride channel complex is known to be the target site of dieldrin, a cyclodiene insecticide. GABA-receptors, with a naturally occurring amino acid substitution, A302S/G in the putative ion-channel lining region, confer resistance to cyclodiene insecticides that includes aldrin, chlordane, dieldrin, heptachlor, endrin and endosulphan. Methods A total of 154 mosquito samples from 10 provinces of malaria-endemic areas across Indonesia (Aceh, North Sumatra, Bangka Belitung, Lampung, Central Java, East Nusa Tenggara, West Nusa Tenggara, West Sulawesi, Molucca and North Molucca) were obtained and identified by species, using morphological characteristic. The DNA was individually extracted using chelex-ion exchanger and the DNA obtained was used for analyses using sequencing method. Results Molecular analysis indicated 11% of the total 154 Anopheles samples examined, carried Rdl mutant alleles. All of the alleles were found in homozygous form. Rdl 302S allele was observed in Anopheles vagus (from Central Java, Lampung, and West Nusa Tenggara), Anopheles aconitus (from Central Java), Anopheles barbirostris (from Central Java and Lampung), Anopheles sundaicus (from North Sumatra and Lampung), Anopheles nigerrimus (from North Sumatra), whereas the 302 G allele was only found in Anopheles farauti from Molucca. Conclusion The existence of the Rdl mutant allele indicates that, either insecticide pressure on the Anopheles population in these areas might still be ongoing (though not directly associated with the malaria control programme) or that the mutant form of the Rdl allele is relatively stable in the absence of insecticide. Nonetheless, the finding suggests that integrated pest management is warranted in malaria-endemic areas where insecticides are widely used for other purposes. PMID:22364613

The resting sites and blood-meal sources of Anopheles minimus in Taiwan

PubMed Central

Chang, Mei-Chun; Teng, Hwa-Jen; Chen, Chen-Fu; Chen, Yung-Chen; Jeng, Chian-Ren

2008-01-01

Background The WHO declared Taiwan free from malaria in 1965, but in 2003 the reporting of two introduced cases in a rural area suggested a possible local transmission of this disease. Therefore, understanding the resting sites and the blood sources of Anopheles minimus is crucial in order to provide information for implementing vector control strategies. Methods During a two-year survey, mosquitoes were collected in houses and their surrounding areas and at the bank of larval habitats by backpack aspirators in 17 villages in rural areas of southern and eastern Taiwan for 1 hr. On the same day, blacklight traps were hung downward overnight. Blood-fed mosquito samples were analysed by PCR. Results Of the 195 total households surveyed by backpack aspirators, no Anopheles adults were collected inside the houses, while a single Anopheles minimus and a single Anopheles maculatus were collected outside of the houses. On the same day, 23 An. minimus, two An. maculatus, two Anopheles ludlowae, two Anopheles sinensis, and one Anopheles tessellatus were collected along the bank of larval habitats. In blacklight traps hung outside of the houses in the villages, 69 An. minimus, 62 An. ludlowae, 31 An. sinensis, and 19 An. maculatus were collected. In larval habitats, 98 An. ludlowae, 64 An. minimus, 49 An. sinensis, and 14 An. maculatus were collected. Of a total of 10 blood-fed samples, An. minimus fed on four animals including bovine (60%), dogs (20%), pig (10%), and non-chicken avian (10%). Conclusion Anopheles minimus, an opportunist feeder in Taiwan, was not collected inside the houses, but was found outside of the houses in villages and surrounding larval habitats. Therefore, an outdoor transmission of malaria is likely to occur and, thus, the bed nets, which are favoured for controlling the late biting of An. minimus, should be a very efficient and effective method for those local residents who sleep outdoors. Additionally, space spray of insecticides for Anopheles at

Desiccation tolerance in Anopheles coluzzii: the effects of spiracle size and cuticular hydrocarbons

PubMed Central

Arcaz, Arthur C.; Huestis, Diana L.; Dao, Adama; Yaro, Alpha S.; Diallo, Moussa; Andersen, John; Blomquist, Gary J.; Lehmann, Tovi

2016-01-01

ABSTRACT The African malaria mosquitoes Anopheles gambiae and Anopheles coluzzii range over forests and arid areas, where they withstand dry spells and months-long dry seasons, suggesting variation in their desiccation tolerance. We subjected a laboratory colony (G3) and wild Sahelian mosquitoes during the rainy and dry seasons to desiccation assays. The thoracic spiracles and amount and composition of cuticular hydrocarbons (CHCs) of individual mosquitoes were measured to determine the effects of these traits on desiccation tolerance. The relative humidity of the assay, body water available, rate of water loss and water content at death accounted for 88% of the variation in desiccation tolerance. Spiracle size did not affect the rate of water loss or desiccation tolerance of the colony mosquitoes, as was the case for the total CHCs. However, six CHCs accounted for 71% of the variation in desiccation tolerance and three accounted for 72% of the variation in the rate of water loss. Wild A. coluzzii exhibited elevated desiccation tolerance during the dry season. During that time, relative thorax and spiracle sizes were smaller than during the rainy season. A smaller spiracle size appeared to increase A. coluzzii's desiccation tolerance, but was not statistically significant. Seasonal changes in CHC composition were detected in Sahelian A. coluzzii. Stepwise regression models suggested the effect of particular CHCs on desiccation tolerance. In conclusion, the combination of particular CHCs along with the total amount of CHCs is a primary mechanism conferring desiccation tolerance in A. coluzzii, while variation in spiracle size might be a secondary mechanism. PMID:27207644

Desiccation tolerance in Anopheles coluzzii: the effects of spiracle size and cuticular hydrocarbons.

PubMed

Arcaz, Arthur C; Huestis, Diana L; Dao, Adama; Yaro, Alpha S; Diallo, Moussa; Andersen, John; Blomquist, Gary J; Lehmann, Tovi

2016-06-01

The African malaria mosquitoes Anopheles gambiae and Anopheles coluzzii range over forests and arid areas, where they withstand dry spells and months-long dry seasons, suggesting variation in their desiccation tolerance. We subjected a laboratory colony (G3) and wild Sahelian mosquitoes during the rainy and dry seasons to desiccation assays. The thoracic spiracles and amount and composition of cuticular hydrocarbons (CHCs) of individual mosquitoes were measured to determine the effects of these traits on desiccation tolerance. The relative humidity of the assay, body water available, rate of water loss and water content at death accounted for 88% of the variation in desiccation tolerance. Spiracle size did not affect the rate of water loss or desiccation tolerance of the colony mosquitoes, as was the case for the total CHCs. However, six CHCs accounted for 71% of the variation in desiccation tolerance and three accounted for 72% of the variation in the rate of water loss. Wild A. coluzzii exhibited elevated desiccation tolerance during the dry season. During that time, relative thorax and spiracle sizes were smaller than during the rainy season. A smaller spiracle size appeared to increase A. coluzzii's desiccation tolerance, but was not statistically significant. Seasonal changes in CHC composition were detected in Sahelian A. coluzzii Stepwise regression models suggested the effect of particular CHCs on desiccation tolerance. In conclusion, the combination of particular CHCs along with the total amount of CHCs is a primary mechanism conferring desiccation tolerance in A. coluzzii, while variation in spiracle size might be a secondary mechanism. © 2016. Published by The Company of Biologists Ltd.

Mass drug administration of ivermectin in south-eastern Senegal reduces the survivorship of wild-caught, blood fed malaria vectors

PubMed Central

2010-01-01

Background In south-eastern Senegal, malaria and onchocerciasis are co-endemic. Onchocerciasis in this region has been controlled by once or twice yearly mass drug administration (MDA) with ivermectin (IVM) for over fifteen years. Since laboratory-raised Anopheles gambiae s.s. are susceptible to ivermectin at concentrations found in human blood post-ingestion of IVM, it is plausible that a similar effect could be quantified in the field, and that IVM might have benefits as a malaria control tool. Methods In 2008 and 2009, wild-caught blood fed An. gambiae s.l. mosquitoes were collected from huts of three pairs of Senegalese villages before and after IVM MDAs. Mosquitoes were held in an insectary to assess their survival rate, subsequently identified to species, and their blood meals were identified. Differences in mosquito survival were statistically analysed using a Glimmix model. Lastly, changes in the daily probability of mosquito survivorship surrounding IVM MDAs were calculated, and these data were inserted into a previously developed, mosquito age-structured model of malaria transmission. Results Anopheles gambiae s.s. (P < 0.0001) and Anopheles arabiensis (P = 0.0191) from the treated villages had significantly reduced survival compared to those from control villages. Furthermore, An gambiae s.s. caught 1-6 days after MDA in treated villages had significantly reduced survival compared to control village collections (P = 0.0003), as well as those caught pre-MDA (P < 0.0001) and >7 days post-MDA (P < 0.0001). The daily probability of mosquito survival dropped >10% for the six days following MDA. The mosquito age-structured model of malaria transmission demonstrated that a single IVM MDA would reduce malaria transmission (Ro) below baseline for at least eleven days, and that repeated IVM MDAs would result in a sustained reduction in malaria Ro. Conclusions Ivermectin MDA significantly reduced the survivorship of An. gambiae s.s. for six days past the date of

Impact of operational effectiveness of long-lasting insecticidal nets (LLINs) on malaria transmission in pyrethroid-resistant areas

PubMed Central

2013-01-01

Background A dynamic study on the transmission of malaria was conducted in two areas (R+ area: Low resistance area; R+++ area: High resistance area) in the department of Plateau in South Eastern Benin, where the population is protected by Long Lasting Insecticidal Nets (LLINs). The aim of this study was to determine if the resistance of malaria vectors to insecticides has an impact on their behavior and on the effectiveness of LLINs in the reduction of malaria transmission. Methods Populations of Anopheles gambiae s.l. were sampled monthly by human landing catch in the two areas to evaluate human biting rates (HBR). Collected mosquitoes were identified morphologically and female Anopheles mosquitoes were tested for the presence of Plasmodium falciparum antigen as assessed using ELISA. The entomological inoculation rate (EIR) was also calculated (EIR = HBR x sporozoitic index [S]). We estimated the parity rate by dissecting the females of An. gambiae. Finally, window catch and spray catch were conducted in order to assess the blood feeding rate and the exophily rate of vectors. Results After 6 months of tracking the mosquito's behavior in contact with the LLINs (Olyset) in R+++ and R+ areas, the entomological indicators of the transmission of malaria (parity rate and sporozoitic index) were similar in the two areas. Also, An. gambiae populations showed the same susceptibility to P. falciparum in both R+ and R+++ areas. The EIR and the exophily rate are higher in R+ area than in R+++ area. But the blood-feeding rate is lower in R+ area comparing to R+++. Conclusion The highest entomological inoculation rate observed in R+ area is mostly due to the strong aggressive density of An. gambiae recorded in one of the study localities. On the other hand, the highest exophily rate and the low blood-feeding rate recorded in R+ area compared to R+++ area are not due to the resistance status of An. gambiae, but due to the differences in distribution and availability of

No Evidence for Lymphatic Filariasis Transmission in Big Cities Affected by Conflict Related Rural-Urban Migration in Sierra Leone and Liberia

PubMed Central

de Souza, Dziedzom K.; Sesay, Santigie; Moore, Marnijina G.; Ansumana, Rashid; Narh, Charles A.; Kollie, Karsor; Rebollo, Maria P.; Koudou, Benjamin G.; Koroma, Joseph B.; Bolay, Fatorma K.; Boakye, Daniel A.; Bockarie, Moses J.

2014-01-01

Background In West Africa, the principal vectors of lymphatic filariasis (LF) are Anopheles species with Culex species playing only a minor role in transmission, if any. Being a predominantly rural disease, the question remains whether conflict-related migration of rural populations into urban areas would be sufficient for active transmission of the parasite. Methodology/Principal Findings We examined LF transmission in urban areas in post-conflict Sierra Leone and Liberia that experienced significant rural-urban migration. Mosquitoes from Freetown and Monrovia, were analyzed for infection with Wuchereria bancrofti. We also undertook a transmission assessment survey (TAS) in Bo and Pujehun districts in Sierra Leone. The majority of the mosquitoes collected were Culex species, while Anopheles species were present in low numbers. The mosquitoes were analyzed in pools, with a maximum of 20 mosquitoes per pool. In both countries, a total of 1731 An. gambiae and 14342 Culex were analyzed for W. bancrofti, using the PCR. Two pools of Culex mosquitoes and 1 pool of An. gambiae were found infected from one community in Freetown. Pool screening analysis indicated a maximum likelihood of infection of 0.004 (95% CI of 0.00012–0.021) and 0.015 (95% CI of 0.0018–0.052) for the An. gambiae and Culex respectively. The results indicate that An. gambiae is present in low numbers, with a microfilaria prevalence breaking threshold value not sufficient to maintain transmission. The results of the TAS in Bo and Pujehun also indicated an antigen prevalence of 0.19% and 0.67% in children, respectively. This is well below the recommended 2% level for stopping MDA in Anopheles transmission areas, according to WHO guidelines. Conclusions We found no evidence for active transmission of LF in cities, where internally displaced persons from rural areas lived for many years during the more than 10 years conflict in Sierra Leone and Liberia. PMID:24516686

Risk factors for house-entry by malaria vectors in a rural town and satellite villages in The Gambia.

PubMed

Kirby, Matthew J; Green, Clare; Milligan, Paul M; Sismanidis, Charalambos; Jasseh, Momadou; Conway, David J; Lindsay, Steven W

2008-01-07

In the pre-intervention year of a randomized controlled trial investigating the protective effects of house screening against malaria-transmitting vectors, a multi-factorial risk factor analysis study was used to identify factors that influence mosquito house entry. Mosquitoes were sampled using CDC light traps in 976 houses, each on one night, in Farafenni town and surrounding villages during the malaria-transmission season in The Gambia. Catches from individual houses were both (a) left unadjusted and (b) adjusted relative to the number of mosquitoes caught in four sentinel houses that were operated nightly throughout the period, to allow for night-to-night variation. Houses were characterized by location, architecture, human occupancy and their mosquito control activities, and the number and type of domestic animals within the compound. 106,536 mosquitoes were caught, of which 55% were Anopheles gambiae sensu lato, the major malaria vectors in the region. There were seven fold higher numbers of An. gambiae s.l. in the villages (geometric mean per trap night = 43.7, 95% confidence intervals, CIs = 39.5-48.4) than in Farafenni town (6.3, 5.7-7.2) and significant variation between residential blocks (p < 0.001). A negative binomial multivariate model performed equally well using unadjusted or adjusted trap data. Using the unadjusted data the presence of nuisance mosquitoes was reduced if the house was located in the town (odds ratio, OR = 0.11, 95% CIs = 0.09-0.13), the eaves were closed (OR = 0.71, 0.60-0.85), a horse was tethered near the house (OR = 0.77, 0.73-0.82), and churai, a local incense, was burned in the room at night (OR = 0.56, 0.47-0.66). Mosquito numbers increased per additional person in the house (OR = 1.04, 1.02-1.06) or trapping room (OR = 1.19, 1.13-1.25) and when the walls were made of mud blocks compared with concrete (OR = 1.44, 1.10-1.87). This study demonstrates that the risk of malaria transmission is greatest in rural areas, where large

Arm-specific dynamics of chromosome evolution in malaria mosquitoes

PubMed Central

2011-01-01

Background The malaria mosquito species of subgenus Cellia have rich inversion polymorphisms that correlate with environmental variables. Polymorphic inversions tend to cluster on the chromosomal arms 2R and 2L but not on X, 3R and 3L in Anopheles gambiae and homologous arms in other species. However, it is unknown whether polymorphic inversions on homologous chromosomal arms of distantly related species from subgenus Cellia nonrandomly share similar sets of genes. It is also unclear if the evolutionary breakage of inversion-poor chromosomal arms is under constraints. Results To gain a better understanding of the arm-specific differences in the rates of genome rearrangements, we compared gene orders and established syntenic relationships among Anopheles gambiae, Anopheles funestus, and Anopheles stephensi. We provided evidence that polymorphic inversions on the 2R arms in these three species nonrandomly captured similar sets of genes. This nonrandom distribution of genes was not only a result of preservation of ancestral gene order but also an outcome of extensive reshuffling of gene orders that created new combinations of homologous genes within independently originated polymorphic inversions. The statistical analysis of distribution of conserved gene orders demonstrated that the autosomal arms differ in their tolerance to generating evolutionary breakpoints. The fastest evolving 2R autosomal arm was enriched with gene blocks conserved between only a pair of species. In contrast, all identified syntenic blocks were preserved on the slowly evolving 3R arm of An. gambiae and on the homologous arms of An. funestus and An. stephensi. Conclusions Our results suggest that natural selection favors specific gene combinations within polymorphic inversions when distant species are exposed to similar environmental pressures. This knowledge could be useful for the discovery of genes responsible for an association of inversion polymorphisms with phenotypic variations in

Efficacy of two different house screening interventions against exposure to malaria and anaemia in children in The Gambia: a randomized controlled trial

PubMed Central

Kirby, Matthew J; Ameh, David; Green, Clare; Jawara, Musa; Milligan, Paul J; Bottomley, Christian; Snell, Paul C; Conway, David J; Lindsay, Steve W

2013-01-01

Summary Background House screening should protect people against malaria. We assessed whether two types of house screening, full screening of windows, doors and closing eaves or installing netting ceilings in local houses, could reduce malaria vector house entry and anaemia in children, in an area of seasonal transmission. Methods 500 occupied houses in and near Farafenni town in The Gambia were randomly assigned to receive full screening, screened ceilings, or no screening, in an area where coverage of insecticide-treated nets was low. Screening was not treated with insecticide. Exposure to mosquitoes indoors was assessed by fortnightly light trap collections, and haemoglobin (Hb) concentration and the prevalence of anaemia and parasitaemia measured in children, aged 6 months to 10 years old, at the end of the transmission season. Findings The mean number of Anopheles gambiae sensu lato mosquitoes, the principal malaria vector, was reduced by 59% in houses with full screening (95% CI 46, 69; p<0.001) and 47% in houses with screened ceilings (95% CI 30, 60; p<0.001) compared with unscreened houses (37.5/trap/night). Anaemia prevalence (Hb <80g/L) was 19.0% among children in unscreened houses, compared to 12.3% with full screening, adjusted odds ratio (OR) 0.53 (0.29, 0.97; p=0.04), and 11.7% with screened ceilings, adjusted OR 0.51, (0.27, 0.96; p=0.04). Mean Hb concentration was higher in children living in fully screened houses and in houses with screened ceilings (104g/L in both groups), than those in unscreened houses (100g/L); adjusted estimates of the differences 3.7g/L (0.3, 7.2; p=0.03) and 4.2g/L (0.6, 7.7; p=0.02) respectively. There was no evidence of an effect on the prevalence of malaria infection. Interpretation House screening substantially reduced the number of mosquitoes inside houses and can contribute to prevention of anaemia in children. Funding Medical Research Council PMID:19732949

Fine structure of the eggs of Anopheles (Anopheles) apicimacula (Diptera:Culicidae).

PubMed

Rodriguez, M H; Chávez, B; Orozco, A; Martínez-Palomo, A

1996-09-01

The eggs of Anopheles (Anopheles) apicimacula Dyar and Knab are described from scanning electron micrographs. The eggs are boat-shaped, with frills that extend ventrally along the length of the egg and surround the deck region. The ornamentation on the dorsal and lateral surfaces is formed by groups of smooth, round tubercles. The ventral surface is covered by irregularly jagged tubercles. Prominent lobed tubercles are present at the anterior and posterior ends of the deck.

Human Antibody Responses to the Anopheles Salivary gSG6-P1 Peptide: A Novel Tool for Evaluating the Efficacy of ITNs in Malaria Vector Control

PubMed Central

Drame, Papa Makhtar; Poinsignon, Anne; Besnard, Patrick; Cornelie, Sylvie; Le Mire, Jacques; Toto, Jean-Claude; Foumane, Vincent; Dos-Santos, Maria Adelaide; Sembène, Mbacké; Fortes, Filomeno; Simondon, Francois; Carnevale, Pierre; Remoue, Franck

2010-01-01

To optimize malaria control, WHO has prioritised the need for new indicators to evaluate the efficacy of malaria vector control strategies. The gSG6-P1 peptide from gSG6 protein of Anopheles gambiae salivary glands was previously designed as a specific salivary sequence of malaria vector species. It was shown that the quantification of human antibody (Ab) responses to Anopheles salivary proteins in general and especially to the gSG6-P1 peptide was a pertinent biomarker of human exposure to Anopheles. The present objective was to validate this indicator in the evaluation of the efficacy of Insecticide Treated Nets (ITNs). A longitudinal evaluation, including parasitological, entomological and immunological assessments, was conducted on children and adults from a malaria-endemic area before and after the introduction of ITNs. Significant decrease of anti-gSG6-P1 IgG response was observed just after the efficient ITNs use. Interestingly, specific IgG Ab level was especially pertinent to evaluate a short-time period of ITNs efficacy and at individual level. However, specific IgG rose back up within four months as correct ITN use waned. IgG responses to one salivary peptide could constitute a reliable biomarker for the evaluation of ITN efficacy, at short- and long-term use, and provide a valuable tool in malaria vector control based on a real measurement of human-vector contact. PMID:21179476

Spatio-temporal distribution of mosquitoes and risk of malaria infection in Rwanda.

PubMed

Hakizimana, Emmanuel; Karema, Corine; Munyakanage, Dunia; Githure, John; Mazarati, Jean Baptiste; Tongren, Jon Eric; Takken, Willem; Binagwaho, Agnes; Koenraadt, Constantianus J M

2018-06-01

To date, the Republic of Rwanda has not systematically reported on distribution, diversity and malaria infectivity rate of mosquito species throughout the country. Therefore, we assessed the spatial and temporal variation of mosquitoes in the domestic environment, as well as the nocturnal biting behavior and infection patterns of the main malaria vectors in Rwanda. For this purpose, mosquitoes were collected monthly from 2010 to 2013 by human landing catches (HLC) and pyrethrum spray collections (PSC) in seven sentinel sites. Mosquitoes were identified using morphological characteristics and PCR. Plasmodium falciparum sporozoite infection rates were determined using ELISA. A total of 340,684 mosquitoes was collected by HLC and 73.8% were morphologically identified as culicines and 26.2% as anophelines. Of the latter, 94.3% were Anopheles gambiae s.l., 0.4% Anopheles funestus and 5.3% other Anopheles species. Of An. gambiae s.l., An. arabiensis and An. gambiae s.s. represented 84.4% and 15.6%, respectively. Of all An. gambiae s.l. collected indoor and outdoor, the proportion collected indoors was 51.3% in 2010 and 44.9% in 2013. A total of 17,022 mosquitoes was collected by PSC of which 20.5% were An. gambiae s.l. and 79.5% were culicines. For the seven sentinel sites, the mean indoor density for An. gambiae s.l. varied from 0.0 to 1.0 mosquitoes/house/night. P. falciparum infection rates in mosquitoes varied from 0.87 to 4.06%. The entomological inoculation rate (EIR) ranged from 1.0 to 329.8 with an annual average of 99.5 infective bites/person/year. This longitudinal study shows, for the first time, the abundance, species composition, and entomological inoculation rate of malaria mosquitoes collected throughout Rwanda. Copyright © 2018 The Authors. Published by Elsevier B.V. All rights reserved.

Seasonal prevalence of malaria vectors and entomological inoculation rates in the rubber cultivated area of Niete, South Region of Cameroon

PubMed Central

2012-01-01

Background Development of large scale agro-industries are subject to serious environmental modifications. In malaria endemic areas this would greatly impact on the transmission paradigm. Two cross-sectional entomological surveys to characterize the Anopheles fauna and their entomological inoculation rates were conducted during May 2010 (peak rainy season) and December 2010 (peak dry season) in the intense rubber cultivated area of Niete in southern forested Cameroon. Methods Mosquitoes were sampled by night collections on human volunteers, identified morphologically and members of the Anopheles gambiae complex further identified to species and molecular form. Parity status was determined following the dissection of the ovaries. Plasmodium falciparum circumsporozoite antigen indices were estimated after the identification of CS antigen by ELISA and the average entomological inoculation rates determined. Results A total of 1187 Anopheles was collected, 419 (35.3%) in the rainy season and 768 (64.7%) in the dry season. Species found were the M molecular form of An. gambiae s.s (66.8%), An. ziemanni (28.3%), An. paludis (4.7%), An. smithii (0.2%). An. gambiae M-form was the principal species in the dry (56.2%) and wet (86.2%) seasons. Average overall entomological inoculation rate for the malaria vectors varied between the dry season (1.09 ib/p/n) and the rainy season (2.30 ib/p/n). Conclusions Malaria transmission in Niete occurs both in the dry and rainy season with the intensities peaking in the dry season. This is unlike previous studies in other areas of southern forested Cameroon where transmission generally peaks in the rainy season. Environmental modifications due to agro-industrial activities might have influenced vector distribution and the dynamics of malaria transmission in this area. This necessitates the possible implementation of control strategies that are related to the eco-geography of the area. PMID:22963986

Insecticidal and sterilizing effect of Olyset Duo®, a permethrin and pyriproxyfen mixture net against pyrethroid-susceptible and -resistant strains of Anopheles gambiae s.s.: a release-recapture assay in experimental huts.

PubMed

Djènontin, Armel; Ahoua Alou, Ludovic P; Koffi, Alphonsine; Zogo, Barnabas; Duarte, Elves; N'Guessan, Raphael; Moiroux, Nicolas; Pennetier, Cédric

2015-01-01

In the context of the widespread distribution of pyrethroid resistance among malaria vectors, we did a release-recapture trial in experimental huts to investigate the insecticidal and sterilizing effects of a novel long-lasting net (LN), Olyset® Duo, incorporating a mixture of permethrin (PER) and the insect growth regulator (IGR), pyri-proxyfen (PPF). An LN containing PPF alone and a classic Olyset® Net were tested in parallel as positive controls. The effect of progressive number of holes (6, 30, or 150) that may accrue in nets over time was simulated. We used two laboratory Anopheles gambiae s.s. strains: the susceptible Kisumu strain and the pyrethroid-resistant VK-Per strain having solely kdr as resistance mechanism. The effect of these nets on the reproductive success of blood-fed females that survived the different LNs conditions was recorded. Regardless of the mosquito strain, the LNs containing PPF alone with as many as 30 holes drastically reduced the number of eggs laid by females succeeding in feeding, i.e. fecundity by 98% and egg hatching rate (fertility) by 93% relative to untreated control net. Very few of the resistant females blood fed and survived under the Olyset® Duo with similar number of holes (up to 30) but of these few, the inhibition of reproductive success was 100%. There was no evidence that the Olyset® Duo LN with 150 holes impacted fecundity or fertility of the resistant colony. The efficacy of Olyset® Duo is encouraging and clearly illustrates that this new net might be a promising tool for malaria transmission control and resistance management. © A. Djènontin et al., published by EDP Sciences, 2015.

Comparison of transmission parameters between Anopheles argyritarsis and Anopheles pseudopunctipennis in two ecologically different localities of Bolivia.

PubMed

Lardeux, Frédéric; Aliaga, Claudia; Tejerina, Rosenka; Torrez, Libia

2013-08-13

Anopheles (Anopheles) pseudopunctipennis is a recognized malaria vector in the slopes of the Andes of Bolivia. There, other species might be involved in malaria transmission and one candidate could be Anopheles argyritarsis. Although it is generally admitted that this species is not a malaria vector in the neotropical region, its potential role in transmission is still controversial and this situation has to be cleared, at least for Bolivia. Comparing the vectorial efficiency of An. pseudopunctipennis with that of An. argyritarsis could solve the question. The two species were sampled throughout Bolivia to estimate their degree of co-existence in their distribution range. Vectorial efficiencies of the two species were compared in two ecologically different localities where the species were sympatric by analysing their vectorial capacities and components (i e, human biting rates, human biting index, survival, durations of the gonotrophic cycle and extrinsic cycle), and the entomological inoculation rates (EIR). Mosquitoes were sampled monthly during more than one year in the two localities. A monthly sample consisted in hourly captures in four houses (inside and outside) in each locality, during four consecutive nights. Climatic variables (temperature, humidity, potential evapo-transpiration and precipitations) were recorded to better understand variability in the entomological parameters. Relationships were analysed using multivariate methods. Anopheles pseudopunctipennis and An. argyritarsis are "altitude" species, sharing the same geographical distribution range in the Andes of Bolivia. No Plasmodium parasite was identified in An. argyritarsis and estimates of the vectorial capacity indicated that it is not a malaria vector in the two studied localities, unlike An. pseudopunctipennis which showed positive EIRs. This latter species, although not a very good malaria vector, exhibited better life traits values and better behavioural characteristics in favour of

Hydrological modeling of geophysical parameters of arboviral and protozoan disease vectors in Internally Displaced People camps in Gulu, Uganda.

PubMed

Jacob, Benjamin G; Muturi, Ephantus J; Caamano, Erick X; Gunter, James T; Mpanga, Enoch; Ayine, Robert; Okelloonen, Joseph; Nyeko, Jack Pen-Mogi; Shililu, Josephat I; Githure, John I; Regens, James L; Novak, Robert J; Kakoma, Ibulaimu

2008-03-14

The aim of this study was to determine if remotely sensed data and Digital Elevation Model (DEM) can test relationships between Culex quinquefasciatus and Anopheles gambiae s.l. larval habitats and environmental parameters within Internally Displaced People (IDP) campgrounds in Gulu, Uganda. A total of 65 georeferenced aquatic habitats in various IDP camps were studied to compare the larval abundance of Cx. quinquefasciatus and An. gambiae s.l. The aquatic habitat dataset were overlaid onto Land Use Land Cover (LULC) maps retrieved from Landsat imagery with 150 m x 150 m grid cells stratified by levels of drainage. The LULC change was estimated over a period of 14 years. Poisson regression analyses and Moran's I statistics were used to model relationships between larval abundance and environmental predictors. Individual larval habitat data were further evaluated in terms of their covariations with spatial autocorrelation by regressing them on candidate spatial filter eigenvectors. Multispectral QuickBird imagery classification and DEM-based GIS methods were generated to evaluate stream flow direction and accumulation for identification of immature Cx. quinquefasciatus and An. gambiae s.l. and abundance. The main LULC change in urban Gulu IDP camps was non-urban to urban, which included about 71.5 % of the land cover. The regression models indicate that counts of An. gambiae s.l. larvae were associated with shade while Cx. quinquefasciatus were associated with floating vegetation. Moran's I and the General G statistics for mosquito density by species and instars, identified significant clusters of high densities of Anopheles; larvae, however, Culex are not consistently clustered. A stepwise negative binomial regression decomposed the immature An. gambiae s.l. data into empirical orthogonal bases. The data suggest the presence of roughly 11% to 28 % redundant information in the larval count samples. The DEM suggest a positive correlation for Culex (0.24) while for

Modeling Malaria Vector Distribution under Climate Change Scenarios in Kenya

NASA Astrophysics Data System (ADS)

Ngaina, J. N.

2017-12-01

Projecting the distribution of malaria vectors under climate change is essential for planning integrated vector control strategies for sustaining elimination and preventing reintroduction of malaria. However, in Kenya, little knowledge exists on the possible effects of climate change on malaria vectors. Here we assess the potential impact of future climate change on locally dominant Anopheles vectors including Anopheles gambiae, Anopheles arabiensis, Anopheles merus, Anopheles funestus, Anopheles pharoensis and Anopheles nili. Environmental data (Climate, Land cover and elevation) and primary empirical geo-located species-presence data were identified. The principle of maximum entropy (Maxent) was used to model the species' potential distribution area under paleoclimate, current and future climates. The Maxent model was highly accurate with a statistically significant AUC value. Simulation-based estimates suggest that the environmentally suitable area (ESA) for Anopheles gambiae, An. arabiensis, An. funestus and An. pharoensis would increase under all two scenarios for mid-century (2016-2045), but decrease for end century (2071-2100). An increase in ESA of An. Funestus was estimated under medium stabilizing (RCP4.5) and very heavy (RCP8.5) emission scenarios for mid-century. Our findings can be applied in various ways such as the identification of additional localities where Anopheles malaria vectors may already exist, but has not yet been detected and the recognition of localities where it is likely to spread to. Moreover, it will help guide future sampling location decisions, help with the planning of vector control suites nationally and encourage broader research inquiry into vector species niche modeling

Finding of Anopheles (Anopheles) hyrcanus (Pallas, 1771) (Diptera, Culicidae) during the entomological surveillance for West Nile virus in Umbria, Italy.

PubMed

Toma, Luciano; Catalani, Monica; Catalano, Antonella; Goffredo, Maria; Romi, Roberto; Di Luca, Marco

2016-08-03

This study reports the mosquito collections conducted in July-August 2013 in Castiglione del Lago (Umbria Region, Italy) during the entomological surveillance within the West Nile national control program. The mosquito collections showed the noteworthy occurrence of Anopheles hyrcanus s.l. (n = 156; 35.8% of the whole sample), a relatively rare mosquito species in Italy, with molecular analyses con rming that these mosquitoes belong to An. hyrcanus s.s. The same catching site was characterized by a relevant richness in mosquito species, in particular the following were found: Anopheles maculipennis s.l. (n = 146; 33.5% composed by 80% Anopheles melanoon, 13.3% Anopheles maculipennis s.s. and 6.7% Anopheles atroparvus), Culex pipiens s.l. (n = 116; 26.6% composed by 90% Cx. p. pipiens, 5% Cx. p. molestus and 5% Cx. p. pipiens/molestus), Culiseta annulata (n = 10; 2.3%), Aedes albopictus (n = 3; 0.7%), and Anopheles claviger (n = 5; 100%).

Green tea proanthocyanidins cause impairment of hormone-regulated larval development and reproductive fitness via repression of juvenile hormone acid methyltransferase, insulin-like peptide and cytochrome P450 genes in Anopheles gambiae sensu stricto.

PubMed

Muema, Jackson M; Nyanjom, Steven G; Mutunga, James M; Njeru, Sospeter N; Bargul, Joel L

2017-01-01

Successful optimization of plant-derived compounds into control of nuisance insects would benefit from scientifically validated targets. However, the close association between the genotypic responses and physiological toxicity effects mediated by these compounds remains underexplored. In this study, we evaluated the sublethal dose effects of proanthocyanidins (PAs) sourced from green tea (Camellia sinensis) on life history traits of Anopheles gambiae (sensu stricto) mosquitoes with an aim to unravel the probable molecular targets. Based on the induced phenotypic effects, genes selected for study targeted juvenile hormone (JH) biosynthesis, signal transduction, oxidative stress response and xenobiotic detoxification in addition to vitellogenesis in females. Our findings suggest that chronic exposure of larval stages (L3/L4) to sublethal dose of 5 ppm dramatically extended larval developmental period for up to 12 days, slowed down pupation rates, induced abnormal larval-pupal intermediates and caused 100% inhibition of adult emergence. Further, females exhibited significant interference of fecundity and egg hatchability relative to controls (p < 0.001). Using reverse transcription quantitative polymerase chain reaction (RT-qPCR), our findings show that PA-treated larvae exhibited significant repression of AgamJHAMT (p < 0.001), AgamILP1 (p < 0.001) and AgamCYP6M2 (p < 0.001) with up-regulation of Hsp70 (p < 0.001). Females exposed as larvae demonstrated down-regulation of AgamVg (p = 0.03), AgamILP1 (p = 0.009), AgamCYP6M2 (p = 0.05) and AgamJHAMT (p = 0.02). Our findings support that C. sinensis proanthocyanidins affect important vectorial capacity components such as mosquito survival rates and reproductive fitness thus could be potentially used for controlling populations of malaria vectors.

Bacterial Diversity Associated with Wild Caught Anopheles Mosquitoes from Dak Nong Province, Vietnam Using Culture and DNA Fingerprint

PubMed Central

Ngo, Chung Thuy; Aujoulat, Fabien; Veas, Francisco; Jumas-Bilak, Estelle; Manguin, Sylvie

2015-01-01

Background Microbiota of Anopheles midgut can modulate vector immunity and block Plasmodium development. Investigation on the bacterial biodiversity in Anopheles, and specifically on the identification of bacteria that might be used in malaria transmission blocking approaches, has been mainly conducted on malaria vectors of Africa. Vietnam is an endemic country for both malaria and Bancroftian filariasis whose parasitic agents can be transmitted by the same Anopheles species. No information on the microbiota of Anopheles mosquitoes in Vietnam was available previous to this study. Method The culture dependent approach, using different mediums, and culture independent (16S rRNA PCR – TTGE) method were used to investigate the bacterial biodiversity in the abdomen of 5 Anopheles species collected from Dak Nong Province, central-south Vietnam. Molecular methods, sequencing and phylogenetic analysis were used to characterize the microbiota. Results and Discussion The microbiota in wild-caught Anopheles was diverse with the presence of 47 bacterial OTUs belonging to 30 genera, including bacterial genera impacting Plasmodium development. The bacteria were affiliated with 4 phyla, Actinobacteria, Bacteroidetes, Firmicutes and Proteobacteria, the latter being the dominant phylum. Four bacterial genera are newly described in Anopheles mosquitoes including Coxiella, Yersinia, Xanthomonas, and Knoellia. The bacterial diversity per specimen was low ranging from 1 to 4. The results show the importance of pairing culture and fingerprint methods to better screen the bacterial community in Anopheles mosquitoes. Conclusion Sampled Anopheles species from central-south Vietnam contained a diverse bacterial microbiota that needs to be investigated further in order to develop new malaria control approaches. The combination of both culture and DNA fingerprint methods allowed a thorough and complementary screening of the bacterial community in Anopheles mosquitoes. PMID:25747513

Interventions that effectively target Anopheles funestus mosquitoes could significantly improve control of persistent malaria transmission in south-eastern Tanzania.

PubMed

Kaindoa, Emmanuel W; Matowo, Nancy S; Ngowo, Halfan S; Mkandawile, Gustav; Mmbando, Arnold; Finda, Marcelina; Okumu, Fredros O

2017-01-01

Malaria is transmitted by many Anopheles species whose proportionate contributions vary across settings. We re-assessed the roles of Anopheles arabiensis and Anopheles funestus, and examined potential benefits of species-specific interventions in an area in south-eastern Tanzania, where malaria transmission persists, four years after mass distribution of long-lasting insecticide-treated nets (LLINs). Monthly mosquito sampling was done in randomly selected households in three villages using CDC light traps and back-pack aspirators, between January-2015 and January-2016, four years after the last mass distribution of LLINs in 2011. Multiplex polymerase chain reaction (PCR) was used to identify members of An. funestus and Anopheles gambiae complexes. Enzyme-linked immunosorbent assay (ELISA) was used to detect Plasmodium sporozoites in mosquito salivary glands, and to identify sources of mosquito blood meals. WHO susceptibility assays were done on wild caught female An. funestus s.l, and physiological ages approximated by examining mosquito ovaries for parity. A total of 20,135 An. arabiensis and 4,759 An. funestus were collected. The An. funestus group consisted of 76.6% An. funestus s.s, 2.9% An. rivulorum, 7.1% An. leesoni, and 13.4% unamplified samples. Of all mosquitoes positive for Plasmodium, 82.6% were An. funestus s.s, 14.0% were An. arabiensis and 3.4% were An. rivulorum. An. funestus and An. arabiensis contributed 86.21% and 13.79% respectively, of annual entomological inoculation rate (EIR). An. arabiensis fed on humans (73.4%), cattle (22.0%), dogs (3.1%) and chicken (1.5%), but An. funestus fed exclusively on humans. The An. funestus populations were 100% susceptible to organophosphates, pirimiphos methyl and malathion, but resistant to permethrin (10.5% mortality), deltamethrin (18.7%), lambda-cyhalothrin (18.7%) and DDT (26.2%), and had reduced susceptibility to bendiocarb (95%) and propoxur (90.1%). Parity rate was higher in An. funestus (65.8%) than

Interventions that effectively target Anopheles funestus mosquitoes could significantly improve control of persistent malaria transmission in south–eastern Tanzania

PubMed Central

Matowo, Nancy S.; Ngowo, Halfan S.; Mkandawile, Gustav; Mmbando, Arnold; Finda, Marcelina; Okumu, Fredros O.

2017-01-01

Malaria is transmitted by many Anopheles species whose proportionate contributions vary across settings. We re-assessed the roles of Anopheles arabiensis and Anopheles funestus, and examined potential benefits of species-specific interventions in an area in south-eastern Tanzania, where malaria transmission persists, four years after mass distribution of long-lasting insecticide-treated nets (LLINs). Monthly mosquito sampling was done in randomly selected households in three villages using CDC light traps and back-pack aspirators, between January-2015 and January-2016, four years after the last mass distribution of LLINs in 2011. Multiplex polymerase chain reaction (PCR) was used to identify members of An. funestus and Anopheles gambiae complexes. Enzyme-linked immunosorbent assay (ELISA) was used to detect Plasmodium sporozoites in mosquito salivary glands, and to identify sources of mosquito blood meals. WHO susceptibility assays were done on wild caught female An. funestus s.l, and physiological ages approximated by examining mosquito ovaries for parity. A total of 20,135 An. arabiensis and 4,759 An. funestus were collected. The An. funestus group consisted of 76.6% An. funestus s.s, 2.9% An. rivulorum, 7.1% An. leesoni, and 13.4% unamplified samples. Of all mosquitoes positive for Plasmodium, 82.6% were An. funestus s.s, 14.0% were An. arabiensis and 3.4% were An. rivulorum. An. funestus and An. arabiensis contributed 86.21% and 13.79% respectively, of annual entomological inoculation rate (EIR). An. arabiensis fed on humans (73.4%), cattle (22.0%), dogs (3.1%) and chicken (1.5%), but An. funestus fed exclusively on humans. The An. funestus populations were 100% susceptible to organophosphates, pirimiphos methyl and malathion, but resistant to permethrin (10.5% mortality), deltamethrin (18.7%), lambda-cyhalothrin (18.7%) and DDT (26.2%), and had reduced susceptibility to bendiocarb (95%) and propoxur (90.1%). Parity rate was higher in An. funestus (65.8%) than

Frequency of multiple blood meals taken in a single gonotrophic cycle by Anopheles arabiensis mosquitoes in Macha, Zambia.

PubMed

Norris, Laura C; Fornadel, Christen M; Hung, Wei-Chien; Pineda, Fernando J; Norris, Douglas E

2010-07-01

Anopheles arabiensis is a major vector of Plasmodium falciparum in southern Zambia. This study aimed to determine the rate of multiple human blood meals taken by An. arabiensis to more accurately estimate entomologic inoculation rates (EIRs). Mosquitoes were collected in four village areas over two seasons. DNA from human blood meals was extracted and amplified at four microsatellite loci. Using the three-allele method, which counts > or = 3 alleles at any microsatellite locus as a multiple blood meal, we determined that the overall frequency of multiple blood meals was 18.9%, which was higher than rates reported for An. gambiae in Kenya and An. funestus in Tanzania. Computer simulations showed that the three-allele method underestimates the true multiple blood meal proportion by 3-5%. Although P. falciparum infection status was not shown to influence the frequency of multiple blood feeding, the high multiple feeding rate found in this study increased predicted malaria risk by increasing EIR.

Situation Report--Australia, The Gambia, Papua and New Guinea, Rhodesia, Sri Lanka, Taiwan, Tanzania, Tonga, and Western Samoa.

ERIC Educational Resources Information Center

International Planned Parenthood Federation, London (England).

Data relating to population and family planning in nine foreign countries are presented in these situation reports. Countries included are Australia, The Gambia, Papua and New Guinea, Rhodesia, Sri Lanka, Taiwan, Tanzania, Tonga, and Western Somoa. Information is provided under three topics, statistical information, general background information,…

Shifts in malaria vector species composition and transmission dynamics along the Kenyan coast over the past 20 years.

PubMed

Mwangangi, Joseph M; Mbogo, Charles M; Orindi, Benedict O; Muturi, Ephantus J; Midega, Janet T; Nzovu, Joseph; Gatakaa, Hellen; Githure, John; Borgemeister, Christian; Keating, Joseph; Beier, John C

2013-01-08

Over the past 20 years, numerous studies have investigated the ecology and behaviour of malaria vectors and Plasmodium falciparum malaria transmission on the coast of Kenya. Substantial progress has been made to control vector populations and reduce high malaria prevalence and severe disease. The goal of this paper was to examine trends over the past 20 years in Anopheles species composition, density, blood-feeding behaviour, and P. falciparum sporozoite transmission along the coast of Kenya. Using data collected from 1990 to 2010, vector density, species composition, blood-feeding patterns, and malaria transmission intensity was examined along the Kenyan coast. Mosquitoes were identified to species, based on morphological characteristics and DNA extracted from Anopheles gambiae for amplification. Using negative binomial generalized estimating equations, mosquito abundance over the period were modelled while adjusting for season. A multiple logistic regression model was used to analyse the sporozoite rates. Results show that in some areas along the Kenyan coast, Anopheles arabiensis and Anopheles merus have replaced An. gambiae sensu stricto (s.s.) and Anopheles funestus as the major mosquito species. Further, there has been a shift from human to animal feeding for both An. gambiae sensu lato (s.l.) (99% to 16%) and An. funestus (100% to 3%), and P. falciparum sporozoite rates have significantly declined over the last 20 years, with the lowest sporozoite rates being observed in 2007 (0.19%) and 2008 (0.34%). There has been, on average, a significant reduction in the abundance of An. gambiae s.l. over the years (IRR = 0.94, 95% CI 0.90-0.98), with the density standing at low levels of an average 0.006 mosquitoes/house in the year 2010. Reductions in the densities of the major malaria vectors and a shift from human to animal feeding have contributed to the decreased burden of malaria along the Kenyan coast. Vector species composition remains heterogeneous but in

Species Composition and Distribution of Adult Anopheles (Diptera: Culicidae) in Panama

PubMed Central

LOAIZA, J. R.; BERMINGHAM, E.; SCOTT, M. E.; ROVIRA, J. R.; CONN, J. E.

2010-01-01

Anopheles (Diptera: Culicidae) species composition and distribution were studied using human landing catch data over a 35-yr period in Panama. Mosquitoes were collected from 77 sites during 228 field trips carried out by members of the National Malaria Eradication Service. Fourteen Anopheles species were identified. The highest average human biting rates were recorded from Anopheles (Nyssorhynchus) albimanus (Wiedemann) (9.8 bites/person/night) and Anopheles (Anopheles) punctimacula (Dyar and Knab) (6.2 bites/person/night). These two species were also the most common, present in 99.1 and 74.9%, respectively, of the sites. Anopheles (Nyssorhynchus) aquasalis (Curry) was encountered mostly in the indigenous Kuna Yala Comarca along the eastern Atlantic coast, where malaria case history and average human biting rate (9.3 bites/person/night) suggest a local role in malaria transmission. An. albimanus, An. punctimacula, and Anopheles (Anopheles) vestitipennis (Dyar and Knab) were more abundant during the rainy season (May–December), whereas An. aquasalis was more abundant in the dry season (January–April). Other vector species collected in this study were Anopheles (Kerteszia) neivai (Howard, Dyar, and Knab) and Anopheles (Anopheles) pseudopunctipennis s.l. (Theobald). High diversity of Anopheles species and six confirmed malaria vectors in endemic areas of Panama emphasize the need for more detailed studies to better understand malaria transmission dynamics. PMID:18826025

Synergy between repellents and organophosphates on bed nets: efficacy and behavioural response of natural free-flying An. gambiae mosquitoes.

PubMed

Pennetier, Cédric; Costantini, Carlo; Corbel, Vincent; Licciardi, Séverine; Dabiré, Roch K; Lapied, Bruno; Chandre, Fabrice; Hougard, Jean-Marc

2009-11-19

Chemicals are used on bed nets in order to prevent infected bites and to kill aggressive malaria vectors. Because pyrethroid resistance has become widespread in the main malaria vectors, research for alternative active ingredients becomes urgent. Mixing a repellent and a non-pyrethroid insecticide seemed to be a promising tool as mixtures in the laboratory showed the same features as pyrethroids. We present here the results of two trials run against free-flying Anopheles gambiae populations comparing the effects of two insect repellents (either DEET or KBR 3023, also known as icaridin) and an organophosphate insecticide at low-doses (pirimiphos-methyl, PM) used alone and in combination on bed nets. We showed that mixtures of PM and the repellents induced higher exophily, blood feeding inhibition and mortality among wild susceptible and resistant malaria vectors than compounds used alone. Nevertheless the synergistic interactions are only involved in the high mortality induced by the two mixtures. These field trials argue in favour of the strategy of mixing repellent and organophosphate on bed nets to better control resistant malaria vectors.

Assessment of Cognitive Abilities in Multiethnic Countries: The Case of the Wolof and Mandinka in the Gambia

ERIC Educational Resources Information Center

Jukes, Matthew C. H.; Grigorenko, Elena L.

2010-01-01

Background: The use of cognitive tests is increasing in Africa but little is known about how such tests are affected by the great ethnic and linguistic diversity on the continent. Aim: To assess ethnic and linguistic group differences in cognitive test performance in the West African country of the Gambia and to investigate the sources of these…

The RooPfs study to assess whether improved housing provides additional protection against clinical malaria over current best practice in The Gambia: study protocol for a randomized controlled study and ancillary studies.

PubMed

Pinder, Margaret; Conteh, Lesong; Jeffries, David; Jones, Caroline; Knudsen, Jakob; Kandeh, Balla; Jawara, Musa; Sicuri, Elisa; D'Alessandro, Umberto; Lindsay, Steve W

2016-06-03

In malaria-endemic areas, residents of modern houses have less malaria than those living in traditional houses. This study will determine if modern housing provides incremental protection against clinical malaria over the current best practice of long-lasting insecticidal nets (LLINs) and prompt treatment in The Gambia, determine the incremental cost-effectiveness of the interventions, and analyze the housing market in The Gambia. A two-armed, household, cluster-randomized, controlled study will be conducted to assess whether improved housing and LLINs combine to provide better protection against clinical malaria in children than LLINs alone in The Gambia. The unit of randomization will be the household, defined as a house and its occupants. A total of 800 households will be enrolled and will receive LLINs, and 400 will receive improved housing before clinical follow-up. One child aged 6 months to 13 years will be enrolled from each household and followed for clinical malaria using active case detection to estimate malaria incidence for two malaria transmission seasons. Episodes of clinical malaria will be the primary endpoint. Study children will be surveyed at the end of each transmission season to estimate the prevalence of Plasmodium falciparum infection, parasite density, and the prevalence of anemia. Exposure to malaria parasites will be assessed using light traps, followed by detection of Anopheles gambiae species and sporozoite infection. Ancillary economic and social science studies will undertake a cost-effectiveness analysis and use qualitative and participatory methods to explore the acceptability of the housing modifications and to design strategies for scaling-up housing interventions. The study is the first of its kind to measure the efficacy of housing on reducing clinical malaria, assess the incremental cost-effectiveness of improved housing, and identify mechanisms for scaling up housing interventions. Trial findings will help inform policy

Combining Technical Competence and Stakeholder Impact in Environmental Education: The Gambia All Schools Nursery Competition

ERIC Educational Resources Information Center

Paulete, Francisca E.; Orr, Blair

2010-01-01

Under the guidance of the Department of Forestry, the Regional Education Directorate, and Peace Corps/The Gambia, the Gambia All Schools Tree Nursery Competition, an environmental education program, was developed to introduce practical environmental education in The Gambia. Data for this report were collected using a rapid appraisal approach.…

Surveillance of malaria vector population density and biting behaviour in western Kenya.

PubMed

Ototo, Ednah N; Mbugi, Jenard P; Wanjala, Christine L; Zhou, Guofa; Githeko, Andrew K; Yan, Guiyun

2015-06-17

Malaria is a great public health burden and Africa suffers the largest share of malaria-attributed deaths. Despite control efforts targeting indoor malaria transmission, such as insecticide-treated bed nets (ITNs) and deployment of indoor residual spraying, transmission of the parasite in western Kenya is still maintained. This study was carried out to determine the impact of ITNs on indoor vector densities and biting behaviour in western Kenya. Indoor collection of adult mosquitoes was done monthly in six study sites in western Kenya using pyrethrum spray collections from 2012 to 2014. The rotator trap collections were done in July-August in 2013 and May-June in 2014. Mosquitoes were collected every 2 h between 18.00 and 08.00 h. Human behaviour study was conducted via questionnaire surveys. Species within Anopheles gambiae complex was differentiated by PCR and sporozoite infectivity was determined by ELISA. Species distribution was determined and bed net coverage in the study sites was recorded. During the study a total of 5,469 mosquito vectors were collected from both PSC and Rotator traps comprising 3,181 (58.2%) Anopheles gambiae and 2,288 (41.8%) Anopheles funestus. Compared to all the study sites, Rae had the highest density of An. gambiae with a mean of 1.2 (P<0.001) while Kombewa had the highest density of An. funestus with a mean of 1.08 (P<0.001). Marani had the lowest density of vectors with 0.06 An. gambiae and 0.17 An. funestus (P<0.001). Among the 700 PCR confirmed An. gambiae s.l. individuals, An. gambiae s.s. accounted for 49% and An. arabiensis 51%. Over 50% of the study population stayed outdoors between 18.00 and 20.00 and 06.00 and 08.00 which was the time when highest densities of blood fed vectors were collected. Anopheles gambie s.s. was the main malaria parasite vector in the highland sites and An. arabiensis in the lowland sites. Bed net ownership in 2012 averaged 87% across the study sites. This study suggests that mass distribution of

A dynamic model of some malaria-transmitting anopheline mosquitoes of the Afrotropical region. I. Model description and sensitivity analysis

PubMed Central

2013-01-01

Background Most of the current biophysical models designed to address the large-scale distribution of malaria assume that transmission of the disease is independent of the vector involved. Another common assumption in these type of model is that the mortality rate of mosquitoes is constant over their life span and that their dispersion is negligible. Mosquito models are important in the prediction of malaria and hence there is a need for a realistic representation of the vectors involved. Results We construct a biophysical model including two competing species, Anopheles gambiae s.s. and Anopheles arabiensis. Sensitivity analysis highlight the importance of relative humidity and mosquito size, the initial conditions and dispersion, and a rarely used parameter, the probability of finding blood. We also show that the assumption of exponential mortality of adult mosquitoes does not match the observed data, and suggest that an age dimension can overcome this problem. Conclusions This study highlights some of the assumptions commonly used when constructing mosquito-malaria models and presents a realistic model of An. gambiae s.s. and An. arabiensis and their interaction. This new mosquito model, OMaWa, can improve our understanding of the dynamics of these vectors, which in turn can be used to understand the dynamics of malaria. PMID:23342980

Traditional use of mosquito-repellent plants in western Kenya and their evaluation in semi-field experimental huts against Anopheles gambiae: ethnobotanical studies and application by thermal expulsion and direct burning.

PubMed

Seyoum, A; Pålsson, K; Kung'a, S; Kabiru, E W; Lwande, W; Killeen, G F; Hassanali, A; Knols, B G J

2002-01-01

Ethnobotanical survey in 2 communities in western Kenya revealed that the most commonly known repellent plants were Ocimum americanum L. (64.1%), Lantana camara L. (17.9%), Tagetes minuta L. (11.3%) and Azadirachta indica A. Juss (8.7%) on Rusinga Island, and Hyptis suaveolens Poit. (49.2%), L. camara (30.9%) and O. basilicum L. (30.4%) in Rambira. Direct burning of plants is the most common method of application for O. americanum (68.8%), L. camara (100%) and O. basilicum (58.8%). Placing branches or whole plants inside houses is most common for H. suaveolens (33.3 and 57.8% for the respective locations), A. indica (66.7 and 100%), and T. minuta (54.8 and 56.0%). The repellency of plants suggested by the ethnobotanical survey and other empirical information was evaluated against the malaria vector Anopheles gambiae s.s. Giles in experimental huts within a screenwalled greenhouse. Thermal expulsion and direct burning were tested as alternative application methods for the selected plants O. americanum, O. kilimandscharicum Guerke, O. suave Willd., L. camara, A. indica, H. suaveolens, Lippia uckambensis Spreng and Corymbia citriodora Hook. When thermally expelled, only H. suaveolens failed to repel mosquitoes, whereas the leaves of C. citriodora (74.5%, P < 0.0001), leaves and seeds of O. suave (53.1%, P < 0.0001) and O. kilimandscharicum (52.0%, P < 0.0001) were the most effective. Leaves of C. citriodora also exhibited the highest repellency (51.3%, P < 0.0001) by direct burning, followed by leaves of L. uckambensis (33.4%, P = 0.0004) and leaves and seeds of O. suave (28.0%, P = 0.0255). The combination of O. kilimandscharicum with L. uckambensis repelled 54.8% of mosquitoes (P < 0.0001) by thermal expulsion. No combination of plants increased repellency by either method. The semi-field system described appears a promising alternative to full-field trials for screening large numbers of candidate repellents without risk of malaria exposure.

Study on the species composition and ecology of anophelines in Addis Zemen, South Gondar, Ethiopia.

PubMed

Kindu, Mizan; Aklilu, Esayas; Balkew, Meshesha; Gebre-Michael, Teshome

2018-03-27

Malaria is a public health problem in Ethiopia and its transmission is generally unstable and seasonal. For the selection of the most appropriate vector control measures, knowledge on the ecology of the vector is necessary at a local level. Therefore, the objectives of this study were to document the species composition, breeding habitat characteristics and occurrence of anopheline larva in Sheni stream and the vectorial role of the prevailing Anopheles in relation to malaria transmission in Addis Zemen, Ethiopia. Immature anophelines were sampled from breeding habitats and characteristics, such as water temperature, turbidity, water current, water pH and other variables, of the habitats were measured from October 2011 to February 2012. Adult anophelines were sampled inside human dwellings using space spray and Center for Disease Control light traps. Artificial pit shelters and clay pots were also used for outdoor adult collections. Anophelines collected were identified using morphological key. The enzyme-linked immunosorbent assay was applied to detect circumsporozoite proteins of Plasmodium and source of blood meals. A total of 6258 Anopheles larvae were collected and identified morphologically. Five anopheline species were found: An. gambiae (s.l.), An. cinereus, An. demeilloni, An. christi and An. pretoriensis. Anopheles gambiae (s.l.) existed in most of the habitats investigated. Only the former three species were captured in the adult collections. Sun-lit Sheni stream, rain pools, hoof prints, drainage and irrigation canals were found to be habitats of larvae. Anopheles gambiae (s.l.) larvae were most abundantly sampled from sand mining and natural sand pools of Sheni stream. Multiple regression analysis showed that clear, permanent and temporary habitats devoid of mats of algae were the best predictors of An. gambiae (s.l.) larval abundance. It is also the responsible malaria vector in the study area and exhibits anthropophilic and endophagic behaviour. The

20-Years of Population-Based Cancer Registration in Hepatitis B and Liver Cancer Prevention in The Gambia, West Africa

PubMed Central

Bah, Ebrima; Carrieri, Maria Patrizia; Hainaut, Pierre; Bah, Yusupha; Nyan, Ousman; Taal, Makie

2013-01-01

Background The Gambia Hepatitis Intervention Study (GHIS) was designed as a randomised control trial of infant hepatitis B vaccination applied to public health policy, with the main goal of preventing primary liver cancer later in adult life in The Gambia. To that effect, the National Cancer Registry of The Gambia (NCR), a population-based cancer registry (PBCR), was established in 1986 to actively collect data on all cancer diagnosis nation-wide. We extracted 20-years (1990-2009) of data to assess for the first time, the evolution of the most common cancers, also describe and demonstrate the role of the PBCR in a hepatitis B and liver cancer prevention programme in this population. Methods and Findings We estimated Age-Standardised Incidence Rates (ASR (W)) of the most common cancers registered during the period by gender. The registration period was divided into four 5-year intervals and incidence rates were estimated for each interval. The most common cancers in males were liver, prostate, lung plus bronchus, non-Hodgkin lymphoma (NHL) and stomach, accounting for 60%, 5%, 4%, 5% and 3%, respectively. Similarly, cancers of the cervix uteri, liver, breast and NHL, were the most common in females, accounting for 33%, 24%, 11% and 4% of the female cancers, respectively. Conclusions Cancer incidence has remained relatively stable over time, but as shown elsewhere in sub-Saharan Africa the disease is a threat in The Gambia. The infection related cancers which are mostly preventable (HBV in men and HPV/HIV in women) were the most common. At the moment the data is not enough to detect an effect of hepatitis B vaccination on liver cancer incidence in The Gambia. However, we observed that monitoring case occurrence through PBCR is a key public health pre-requisite for rational planning and implementation of targeted interventions for improving the health of the population. PMID:24098724

Host attraction and biting behaviour of Anopheles mosquitoes in South Halmahera, Indonesia.

PubMed

St Laurent, Brandyce; Burton, Timothy A; Zubaidah, Siti; Miller, Helen C; Asih, Puji B; Baharuddin, Amirullah; Kosasih, Sully; Shinta; Firman, Saya; Hawley, William A; Burkot, Thomas R; Syafruddin, Din; Sukowati, Supratman; Collins, Frank H; Lobo, Neil F

2017-08-02

Indonesia is home to a variety of malaria vectors whose specific bionomic traits remain largely uncharacterized. Species-specific behaviours, such as host feeding preferences, impact the dynamics of malaria transmission and the effectiveness of vector control interventions. To examine species-specific host attraction and feeding behaviours, a Latin square design was used to compare Anopheles mosquitoes attracted to human, cow, and goat-baited tents. Anopheles mosquitoes were collected hourly from the inside walls of each baited tent. Species were morphologically and then molecularly identified using rDNA ITS2 sequences. The head and thorax of individual specimens were analysed for Plasmodium DNA using PCR. Bloodmeals were identified using a multiplex PCR. A total of 1024, 137, and 74 Anopheles were collected over 12 nights in cow, goat, and human-baited tents, respectively. The species were identified as Anopheles kochi, Anopheles farauti s.s., Anopheles hackeri, Anopheles hinesorum, Anopheles indefinitus, Anopheles punctulatus, Anopheles tessellatus, Anopheles vagus, and Anopheles vanus, many of which are known to transmit human malaria. Molecular analysis of blood meals revealed a high level of feeding on multiple host species in a single night. Anopheles kochi, An. indefinitus, and An. vanus were infected with Plasmodium vivax at rates comparable to primary malaria vectors. The species distributions of Anopheles mosquitoes attracted to human, goat, and cow hosts were similar. Eight of nine sporozoite positive samples were captured with animal-baited traps, indicating that even predominantly zoophilic mosquitoes may be contributing to malaria transmission. Multiple host feeding and flexibility in blood feeding behaviour have important implications for malaria transmission, malaria control, and the effectiveness of intervention and monitoring methods, particularly those that target human-feeding vectors.

Alternative Treatments for Indoor Residual Spraying for Malaria Control in a Village with Pyrethroid- and DDT-Resistant Vectors in The Gambia

PubMed Central

Tangena, Julie-Anne A.; Adiamoh, Majidah; D’Alessandro, Umberto; Jarju, Lamin; Jawara, Musa; Jeffries, David; Malik, Naiela; Nwakanma, Davis; Kaur, Harparkash; Takken, Willem; Lindsay, Steve W.; Pinder, Margaret

2013-01-01

Background Malaria vector control is threatened by resistance to pyrethroids, the only class of insecticides used for treating bed nets. The second major vector control method is indoor residual spraying with pyrethroids or the organochloride DDT. However, resistance to pyrethroids frequently confers resistance to DDT. Therefore, alternative insecticides are urgently needed. Methodology/Principal Findings Insecticide resistance and the efficacy of indoor residual spraying with different insecticides was determined in a Gambian village. Resistance of local vectors to pyrethroids and DDT was high (31% and 46% mortality, respectively) while resistance to bendiocarb and pirimiphos methyl was low (88% and 100% mortality, respectively). The vectors were predominantly Anopheles gambiae s.s. with 94% of them having the putative resistant genotype kdr 1014F. Four groups of eight residential compounds were each sprayed with either (1) bendiocarb, a carbamate, (2) DDT, an organochlorine, (3) microencapsulated pirimiphos methyl, an organophosphate, or (4) left unsprayed. All insecticides tested showed high residual activity up to five months after application. Mosquito house entry, estimated by light traps, was similar in all houses with metal roofs, but was significantly less in IRS houses with thatched roofs (p=0.02). Residents participating in focus group discussions indicated that IRS was considered a necessary nuisance and also may decrease the use of long-lasting insecticidal nets. Conclusion/Significance Bendiocarb and microencapsulated pirimiphos methyl are viable alternatives for indoor residual spraying where resistance to pyrethroids and DDT is high and may assist in the management of pyrethroid resistance. PMID:24058551

PEANUT SHELL FUEL FOR THE GAMBIA

EPA Science Inventory

The project will develop a household-scale human-powered briquette maker that will convert peanut shells into an efficient cooking fuel. The briquette maker will be designed such that it can be manufactured and used in The Gambia.

Phylogenetic study of six species of Anopheles mosquitoes in Peninsular Malaysia based on inter-transcribed spacer region 2 (ITS2) of ribosomal DNA

PubMed Central

2014-01-01

Background Molecular techniques are invaluable for investigation on the biodiversity of Anopheles mosquitoes. This study aimed at investigating the spatial-genetic variations among Anopheles mosquitoes from different areas of Peninsular Malaysia, as well as deciphering evolutionary relationships of the local Anopheles mosquitoes with the mosquitoes from neighbouring countries using the anopheline ITS2 rDNA gene. Methods Mosquitoes were collected, identified, dissected to check infection status, and DNA extraction was performed for PCR with primers targeting the ITS2 rDNA region. Sequencing was done and phylogenetic tree was constructed to study the evolutionary relationship among Anopheles mosquitoes within Peninsular Malaysia, as well as across the Asian region. Results A total of 133 Anopheles mosquitoes consisting of six different species were collected from eight different locations across Peninsular Malaysia. Of these, 65 ITS2 rDNA sequences were obtained. The ITS2 rDNA amplicons of the studied species were of different sizes. One collected species, Anopheles sinensis, shows two distinct pools of population in Peninsular Malaysia, suggesting evolvement of geographic race or allopatric speciation. Conclusion Anopheles mosquitoes from Peninsular Malaysia show close evolutionary relationship with the Asian anophelines. Nevertheless, genetic differences due to geographical segregation can be seen. Meanwhile, some Anopheles mosquitoes in Peninsular Malaysia show vicariance, exemplified by the emergence of distinct cluster of An. sinensis population. PMID:24993022

Organization and startup of The Gambia's new community-based medical programme.

PubMed

Chávez, José A; Suárez, Lázaro V; Del Rosario, Odalis; Hechavarría, Suiberto; Quiñones, Judith

2012-01-01

The shortage of health professionals in developing countries and especially in their poorest regions imperils the vision of health for all. New training policies and strategies are needed urgently to address these shortages. The Gambia's new Community-Based Medical Programme is one such strategy. KEYWORDS Medical education, access to health care, healthcare disparities, health manpower, rural health, developing countries, The Gambia.

Species composition, larval habitats, seasonal occurrence and distribution of potential malaria vectors and associated species of Anopheles (Diptera: Culicidae) from the Republic of Korea

PubMed Central

2010-01-01

Background Larval mosquito habitats of potential malaria vectors and related species of Anopheles from three provinces (Gyeonggi, Gyeongsangbuk, Chungcheongbuk Provinces) of the Republic of Korea were surveyed in 2007. This study aimed to determine the species composition, seasonal occurrence and distributions of Anopheles mosquitoes. Satellite derived normalized difference vegetation index data (NDVI) was also used to study the seasonal abundance patterns of Anopheles mosquitoes. Methods Mosquito larvae from various habitats were collected using a standard larval dipper or a white plastic larval tray, placed in plastic bags, and were preserved in 100% ethyl alcohol for species identification by PCR and DNA sequencing. The habitats in the monthly larval surveys included artificial containers, ground depressions, irrigation ditches, drainage ditches, ground pools, ponds, rice paddies, stream margins, inlets and pools, swamps, and uncultivated fields. All field-collected specimens were identified to species, and relationships among habitats and locations based on species composition were determined using cluster statistical analysis. Results In about 10,000 specimens collected, eight species of Anopheles belonging to three groups were identified: Hyrcanus Group - Anopheles sinensis, Anopheles kleini, Anopheles belenrae, Anopheles pullus, Anopheles lesteri, Anopheles sineroides; Barbirostris Group - Anopheles koreicus; and Lindesayi Group - Anopheles lindesayi japonicus. Only An. sinensis was collected from all habitats groups, while An. kleini, An. pullus and An. sineroides were sampled from all, except artificial containers. The highest number of Anopheles larvae was found in the rice paddies (34.8%), followed by irrigation ditches (23.4%), ponds (17.0%), and stream margins, inlets and pools (12.0%). Anopheles sinensis was the dominant species, followed by An. kleini, An. pullus and An. sineroides. The monthly abundance data of the Anopheles species from three

Ecological Suitability and Spatial Distribution of Five Anopheles Species in Amazonian Brazil

PubMed Central

McKeon, Sascha N.; Schlichting, Carl D.; Povoa, Marinete M.; Conn, Jan E.

2013-01-01

Seventy-six sites characterized in Amazonian Brazil revealed distinct habitat diversification by examining the environmental factors associated with the distribution and abundance of five anopheline species (Diptera: Culicidae) in the subgenus Nyssorhynchus. These included three members of the Albitarsis Complex, Anopheles oryzalimnetes, Anopheles marajoara, Anopheles janconnae; Anopheles triannulatus, and Anopheles goeldii. Anopheles janconnae abundance had a positive correlation to water flow and a negative relationship to sun exposure. Abundance of An. oryzalimentes was associated with water chemistry. Anopheles goeldii larvae were abundant in shaded, more saline waters. Anopheles marajoara and An. triannulatus were negatively associated with available resources, although An. marajoara also showed several local correlations. These analyses suggest An. triannulatus is a habitat generalist, An. oryzalimentes and An. janconnae are specialists, and An. marajoara and An. goeldii could not be easily classified either way. Correlations described herein provide testable hypotheses for future research and identifying habitats for vector control. PMID:23546804

Anophelism in a Former Malaria Area of Northeastern Spain

PubMed Central

Bueno-Marí, Rubén; Jiménez-Peydró, Ricardo

2013-01-01

Background: A field study on diversity and distribution of anophelines currently present in a past endemic malaria area of Spain was carried out in order to identify possible risk areas of local disease transmission. Methods: Multiple larval sites were sampled from June to October of 2011 in the Region of Somontano de Barbastro (Northeastern Spain). The sampling effort was fixed at 10 minutes which included the active search for larvae in each biotope visited. Results: A total of 237 larval specimens belonging to four Anopheles species (Anopheles atroparvus, An. claviger, An. maculipennis and An. petragnani) were collected and identified. Conclusions: Malaria receptivity in the study area is high, especially in the area of Cinca river valley, due to the abundance of breeding sites of An. atroparvus very close to human settlements. Although current socio-economic conditions in Spain reduce possibilities of re-emergence of malaria transmission, it is evident that certain entomological and epidemiological vigilance must be maintained and even increased in the context of current processes of climate change and globalization. PMID:24409440

Pyrethroid resistance in the major malaria vector Anopheles arabiensis from Gwave, a malaria-endemic area in Zimbabwe.

PubMed

Munhenga, Givemore; Masendu, Hieronymo T; Brooke, Basil D; Hunt, Richard H; Koekemoer, Lizette K

2008-11-28

Insecticide resistance can present a major obstacle to malaria control programmes. Following the recent detection of DDT resistance in Anopheles arabiensis in Gokwe, Zimbabwe, the underlying resistance mechanisms in this population were studied. Standard WHO bioassays, using 0.75% permethrin, 4% DDT, 5% malathion, 0.1% bendiocarb and 4% dieldrin were performed on wild-collected adult anopheline mosquitoes and F1 progeny of An. arabiensis reared from wild-caught females. Molecular techniques were used for species identification as well as to identify knockdown resistance (kdr) and ace-1 mutations in individual mosquitoes. Biochemical assays were used to determine the relative levels of detoxifying enzyme systems including non-specific esterases, monooxygenases and glutathione-S-transferases as well as to detect the presence of an altered acetylcholine esterase (AChE). Anopheles arabiensis was the predominant member of the Anopheles gambiae complex. Of the 436 An. arabiensis females, 0.5% were positive for Plasmodium falciparum infection. WHO diagnostic tests on wild populations showed resistance to the pyrethroid insecticide permethrin at a mean mortality of 47% during February 2006 and a mean mortality of 68.2% in January 2008. DDT resistance (68.4% mean mortality) was present in February 2006; however, two years later the mean mortality was 96%. Insecticide susceptibility tests on F1 An. arabiensis families reared from material from two separate collections showed an average mean mortality of 87% (n = 758) after exposure to 4% DDT and 65% (n = 587) after exposure to 0.75% permethrin. Eight families were resistant to both DDT and permethrin. Biochemical analysis of F1 families reared from collections done in 2006 revealed high activity levels of monooxygenase (48.5% of families tested, n = 33, p < 0.05), glutathione S-transferase (25.8% of families tested, n = 31, p < 0.05) and general esterase activity compared to a reference susceptible An. arabiensis colony

Investigating knockdown resistance (kdr) mechanism against pyrethroids/DDT in the malaria vector Anopheles funestus across Africa.

PubMed

Irving, Helen; Wondji, Charles S

2017-08-09

Understanding the molecular basis of insecticide resistance is key to improve the surveillance and monitoring of malaria vector populations under control. In the major malaria vector Anopheles funestus, little is currently known about the role of the knockdown resistance (kdr) mechanism. Here, we investigated the presence and contribution of knockdown resistance (kdr) to pyrethroids/DDT resistance observed in Anopheles funestus across Africa. Pyrosequencing genotyping and sequencing of the voltage gated sodium channel (VGSC) gene did not detect the common L1014F mutation in field collected An. funestus across Africa. Amplification and cloning of the full-length of the sodium channel gene in pyrethroid resistant mosquitoes revealed evidences of alternative splicing events with three transcripts of 2092, 2061 and 2117 amino acids (93% average similarity to An. gambiae). Several amino acid changes were detected close to the domain II of the protein such as L928R, F938 W, I939S, L802S and T1008 M. However, all these mutations are found at low frequency and their role in pyrethroid resistance could not be established. The presence of the exclusive alternative splicing at exon 19 was not associated with resistance phenotype. Analysis of patterns of genetic diversity of the VGSC gene revealed a high polymorphism level of this gene across Africa with no evidence of directional selection suggesting a limited role for knockdown resistance in pyrethroid resistance in An. funestus. Patterns of genetic differentiation correlate with previous observations of the existence of barriers to gene flow Africa-wide with southern population significantly differentiated from other regions. Despite an apparent limited role of knockdown resistance in An. funestus, it is necessary to continue to monitor the contribution of the mutations detected here as increasing selection from insecticide-based interventions may change the dynamic in field populations as previously observed in other

Examining the impact of larval source management and insecticide-treated nets using a spatial agent-based model of Anopheles gambiae and a landscape generator tool.

PubMed

Arifin, S M Niaz; Madey, Gregory R; Collins, Frank H

2013-08-21

Agent-based models (ABMs) have been used to estimate the effects of malaria-control interventions. Early studies have shown the efficacy of larval source management (LSM) and insecticide-treated nets (ITNs) as vector-control interventions, applied both in isolation and in combination. However, the robustness of results can be affected by several important modelling assumptions, including the type of boundary used for landscapes, and the number of replicated simulation runs reported in results. Selection of the ITN coverage definition may also affect the predictive findings. Hence, by replication, independent verification of prior findings of published models bears special importance. A spatially-explicit entomological ABM of Anopheles gambiae is used to simulate the resource-seeking process of mosquitoes in grid-based landscapes. To explore LSM and replicate results of an earlier LSM study, the original landscapes and scenarios are replicated by using a landscape generator tool, and 1,800 replicated simulations are run using absorbing and non-absorbing boundaries. To explore ITNs and evaluate the relative impacts of the different ITN coverage schemes, the settings of an earlier ITN study are replicated, the coverage schemes are defined and simulated, and 9,000 replicated simulations for three ITN parameters (coverage, repellence and mortality) are run. To evaluate LSM and ITNs in combination, landscapes with varying densities of houses and human populations are generated, and 12,000 simulations are run. General agreement with an earlier LSM study is observed when an absorbing boundary is used. However, using a non-absorbing boundary produces significantly different results, which may be attributed to the unrealistic killing effect of an absorbing boundary. Abundance cannot be completely suppressed by removing aquatic habitats within 300 m of houses. Also, with density-dependent oviposition, removal of insufficient number of aquatic habitats may prove counter

Mathematical evaluation of community level impact of combining bed nets and indoor residual spraying upon malaria transmission in areas where the main vectors are Anopheles arabiensis mosquitoes

PubMed Central

2013-01-01

Background Indoor residual insecticide spraying (IRS) and long-lasting insecticide treated nets (LLINs) are commonly used together even though evidence that such combinations confer greater protection against malaria than either method alone is inconsistent. Methods A deterministic model of mosquito life cycle processes was adapted to allow parameterization with results from experimental hut trials of various combinations of untreated nets or LLINs (Olyset®, PermaNet 2.0®, Icon Life® nets) with IRS (pirimiphos methyl, lambda cyhalothrin, DDT), in a setting where vector populations are dominated by Anopheles arabiensis, so that community level impact upon malaria transmission at high coverage could be predicted. Results Intact untreated nets alone provide equivalent personal protection to all three LLINs. Relative to IRS plus untreated nets, community level protection is slightly higher when Olyset® or PermaNet 2.0® nets are added onto IRS with pirimiphos methyl or lambda cyhalothrin but not DDT, and when Icon Life® nets supplement any of the IRS insecticides. Adding IRS onto any net modestly enhances communal protection when pirimiphos methyl is sprayed, while spraying lambda cyhalothrin enhances protection for untreated nets but not LLINs. Addition of DDT reduces communal protection when added to LLINs. Conclusions Where transmission is mediated primarily by An. arabiensis, adding IRS to high LLIN coverage provides only modest incremental benefit (e.g. when an organophosphate like pirimiphos methyl is used), but can be redundant (e.g. when a pyrethroid like lambda cyhalothin is used) or even regressive (e.g. when DDT is used for the IRS). Relative to IRS plus untreated nets, supplementing IRS with LLINs will only modestly improve community protection. Beyond the physical protection that intact nets provide, additional protection against transmission by An. arabiensis conferred by insecticides will be remarkably small, regardless of whether they are delivered

Oviposition Deterrence Induced by Ocimum kilimandscharicum and Ocimum suave Extracts to Gravid Anopheles gambiae s.s (Diptera: Culicidae) in Laboratory

PubMed Central

Kweka, Eliningaya J; Lyatuu, Ester E; Mboya, Michael A; Mwang’onde, Beda J; Mahande, Aneth M

2010-01-01

Background: In most of the past decades, mosquito control has been done by the use of indoor residual spray and insecticides-treated bed nets. The control of mosquitoes by targeting the breeding sites (larval habitat) has not been given priority. Disrupting the oviposition sensory detection of mosquitoes by introducing deterrents of plant origin, which are cheap resources, might be add value to integrated vector control. Such knowledge is required in order to successfully manipulate the behavior of mosquitoes for monitoring or control. Materials and Methods: Twenty gravid mosquitoes were placed in a cage measuring 30 × 30 × 30 cm for oviposition. The oviposition media were made of different materials. Experiments were set up at 6:00 pm, and eggs were collected for counting at 7:30 am. Mosquitoes were observed until they died. The comparisons of the number of eggs were made between the different treatments. Results: There was significant difference in the number of eggs found in control cups when compared with the number of eggs found in water treated with Ocimum kilimandscharicum (OK) (P=0.02) or Ocimum suave (OS) (P=0.000) and that found in water with debris treated with OK (P=0.011) or OS (P=0.002). There was no significant difference in the number of eggs laid in treated water and the number of eggs laid in water with debris treated either with OK (P=0.105) or OS (P=0.176). Oviposition activity index for both OS and OK experiments lay in a negative side and ranged from -0.19% to -1%. The results show that OS and OK deter oviposition in An.gambiae s.s. Conclusions: Further research needs to be done on the effect of secondary metabolites of these plant extracts as they decompose in the breeding sites. In the event of favorable results, the potential of these plant extracts can be harnessed on a larger scale. PMID:20927285

The Gambia and Bangladesh: the seasons and diarrhoea.

PubMed

Rowland, M G

1986-09-01

Climactic factors in the Gambia and Bangladesh have an important impact on the incidence of diarrheal disease. Both countries share some common characteristics in climate, including a cool dry winter of 3 months followed by a hot dry spring and hot wet summers of 5-7 months in length. The main difference is in the amount of rainfall. The Gambia may have 20-30 inches of rain each year; Bangladesh usually has up to 4-5 times this amount. In the Gambia, drought is a recurring problem; floods is the problem in Bangladesh. A study in the Gambia found a close link between the time of the annual peak in diarrhea in young children and the summer rains. A 2nd peak of diarrhea in the winter also was significant and was shown to coincide with a short period of intense transmission of rotavirus. Of the enteric infections of childhood, the enterotoxigenic "Escherichia coli" (ETEC), that is those producing heat-stable toxin (ST) were found to be the most important etiological agents of diarrhea in both countries, with a peak during the rains. In rural Gambia, water is obtained almost exclusively from surface wells, 15-20 meters deep. It was found that, although this water was fecally contaminated throughout the year, levels of contamination increased by up to 100 times with 1-2 days of the start of the rains because excreta is washed into the wells. It also was clear that contaminated water and domestic environment contribute to contamination of children's food. The high level of contamination of food during the summer coincided with the time of high diarrhea prevalence. In Bangladesh it was shown that the incidence of ETEC diarrhea in infants was positively correlated with the frequency of consumption of weaning foods contaminated with fecal coliforms. The seasonal peak of ETEC diarrhea coincided with the time when food was most contaminated due to higher bacterial growth caused by high temperatures. Cholera is endemic in many areas of Bangladesh but not in the Gambia. Though

How effective has tobacco tax increase been in the Gambia? A case study of tobacco control

PubMed Central

Nargis, Nigar; Manneh, Yahya; Krubally, Bakary; Jobe, Baboucarr; Ouma, Ahmed E Ogwell; Tcha-Kondor, Noureiny; Blecher, Evan H

2016-01-01

Objectives The objective of the present study was to evaluate how effective tobacco tax increase has been in increasing price of tobacco products and reducing tobacco consumption in the Gambia. In addition, it tests the hypothesis that tobacco tax revenue grows while tobacco consumption decreases as a result of tax and price increase. Setting The study is designed at the macroeconomic level to examine the import of tobacco products and revenue collected from tobacco taxation in a low-income setting. Participants The participants of this study are the government officials employed in the Ministry of Finance and Economic Affairs (MoFEA), the Gambia and the Gambia Revenue Authority, who are in charge of planning and implementing the tobacco tax policy in the Gambia. Interventions The study includes 2 consecutive interventions in tobacco tax policy in the Gambia. The first intervention was moving the tax base for the uniform specific excise tax on cigarettes from weight to pack of cigarettes in 2013. The second intervention involved increasing the excise and the environmental tax on tobacco products in 2014. Primary and secondary outcome measures The primary outcome measures were the cost, insurance and freight value and the price of tobacco products. The secondary outcome measures included the import of tobacco products and tobacco tax revenue. Results In 2013–2014, the Gambia MoFEA raised the specific excise rate, which increased price, reduced consumption and generated significantly more government revenue from tobacco products. This is a clear evidence of the win-win outcome of raising tobacco tax. In addition, the Gambia has set the example of harmonising tax rates between tobacco products that reduces the substitution between tobacco products. Conclusions The Gambia presents the best practice in tobacco taxation. There is need for documenting more country-specific evidence on the win-win outcome of raising tobacco tax. PMID:27566626

Highly focused anopheline breeding sites and malaria transmission in Dakar

PubMed Central

Machault, Vanessa; Gadiaga, Libasse; Vignolles, Cécile; Jarjaval, Fanny; Bouzid, Samia; Sokhna, Cheikh; Lacaux, Jean-Pierre; Trape, Jean-François; Rogier, Christophe; Pagès, Frédéric

2009-01-01

Background Urbanization has a great impact on the composition of the vector system and malaria transmission dynamics. In Dakar, some malaria cases are autochthonous but parasite rates and incidences of clinical malaria attacks have been recorded at low levels. Ecological heterogeneity of malaria transmission was investigated in Dakar, in order to characterize the Anopheles breeding sites in the city and to study the dynamics of larval density and adult aggressiveness in ten characteristically different urban areas. Methods Ten study areas were sampled in Dakar and Pikine. Mosquitoes were collected by human landing collection during four nights in each area (120 person-nights). The Plasmodium falciparum circumsporozoite (CSP) index was measured by ELISA and the entomological inoculation rates (EIR) were calculated. Open water collections in the study areas were monitored weekly for physico-chemical characterization and the presence of anopheline larvae. Adult mosquitoes and hatched larvae were identified morphologically and by molecular methods. Results In September-October 2007, 19,451 adult mosquitoes were caught among which, 1,101 were Anopheles gambiae s.l. The Human Biting Rate ranged from 0.1 bites per person per night in Yoff Village to 43.7 in Almadies. Seven out of 1,101 An. gambiae s.l. were found to be positive for P. falciparum (CSP index = 0.64%). EIR ranged from 0 infected bites per person per year in Yoff Village to 16.8 in Almadies. The An. gambiae complex population was composed of Anopheles arabiensis (94.8%) and Anopheles melas (5.2%). None of the An. melas were infected with P. falciparum. Of the 54 water collection sites monitored, 33 (61.1%) served as anopheline breeding sites on at least one observation. No An. melas was identified among the larval samples. Some physico-chemical characteristics of water bodies were associated with the presence/absence of anopheline larvae and with larval density. A very close parallel between larval and adult

Household-level and surrounding peri-domestic environmental characteristics associated with malaria vectors Anopheles arabiensis and Anopheles funestus along an urban-rural continuum in Blantyre, Malawi.

PubMed

Dear, Nicole F; Kadangwe, Chifundo; Mzilahowa, Themba; Bauleni, Andy; Mathanga, Don P; Duster, Chifundo; Walker, Edward D; Wilson, Mark L

2018-06-08

Malaria is increasing in some recently urbanized areas that historically were considered lower risk. Understanding what drives urban transmission is hampered by inconsistencies in how "urban" contexts are defined. A dichotomized "urban-rural" approach, based on political boundaries may misclassify environments or fail to capture local drivers of risk. Small-scale agriculture in urban or peri-urban settings has been shown to be a major risk determinant. Household-level Anopheles abundance patterns in and around Malawi's commercial capital of Blantyre (~ 1.9 M pop.) were analysed. Clusters (N = 64) of five houses each located at 2.5 km intervals along eight transects radiating out from Blantyre city centre were sampled during rainy and dry seasons of 2015 and 2016. Mosquito densities were measured inside houses using aspirators to sample resting mosquitoes, and un-baited CDC light traps to sample host seeking mosquitoes. Of 38,895 mosquitoes captured, 91% were female and 87% were Culex spp. Anopheles females (N = 5058) were primarily captured in light traps (97%). Anopheles abundance was greater during rainy seasons. Anopheles funestus was more abundant than Anopheles arabiensis, but both were found on all transects, and had similar associations with environmental risk factors. Anopheles funestus and An. arabiensis females significantly increased with distance from the urban centre, but this trend was not consistent across all transects. Presence of small-scale agriculture was predictive of greater Anopheles spp. abundance, even after controlling for urbanicity, number of nets per person, number of under-5-year olds, years of education, and season. This study revealed how small-scale agriculture along a rural-to-urban transition was associated with An. arabiensis and An. funestus indoor abundances, and that indoor Anopheles density can be high within Blantyre city limits, particularly where agriculture is present. Typical rural areas with lower house

Indoor residual spraying with a mixture of clothianidin (a neonicotinoid insecticide) and deltamethrin provides improved control and long residual activity against pyrethroid resistant Anopheles gambiae sl in Southern Benin.

PubMed

Ngufor, Corine; Fongnikin, Augustin; Rowland, Mark; N'Guessan, Raphael

2017-01-01

There is an urgent need for new insecticides for indoor residual spraying (IRS) which can provide improved and prolonged control of malaria vectors that have developed resistance to existing insecticides. The neonicotinoid, clothianidin represents a class of chemistry new to public health. Clothianidin acts as an agonist on nicotinic acetyl choline receptors. IRS with a mixture of Clothianidin and another WHO approved insecticide such as deltamethrin could provide improved control of insecticide resistant malaria vector populations and serve as a tool for insecticide resistance management. The efficacy and residual activity of a novel IRS mixture of deltamethrin and clothianidin was evaluated against wild pyrethroid resistant An. gambiae sl in experimental huts in Cove, Benin. Two application rates of the mixture were tested and comparison was made with clothianidin and deltamethrin applied alone. To assess the residual efficacy of the treatments on different local wall substrates, the inner walls of the experimental huts were covered with either cement, mud or plywood. Clothianidin demonstrated a clear delayed expression in mortality of wild pyrethroid resistant An. gambiae sl in the experimental huts which reached its full effect 120 hours after exposure. Overall mortality over the 12-month hut trial was 15% in the control hut and 24-29% in the deltamethrin-treated huts. The mixture of clothianidin 200mg/m2 and deltamethrin 25mg/m2 induced high overall hut mortality rates (87% on mud walls, 82% on cement walls and 61% on wooden walls) largely due to the clothianidin component and high hut exiting rates (67-76%) mostly due to the deltamethrin component. Mortality rates remained >80% for 8-9 months on mud and cement walls. The residual activity trend was confirmed by results from monthly in situ cone bioassays with laboratory susceptible An. gambiae Kisumu strain. IRS campaigns with the mixture of clothianidin plus deltamethrin have the potential to provide

Indoor residual spraying with a mixture of clothianidin (a neonicotinoid insecticide) and deltamethrin provides improved control and long residual activity against pyrethroid resistant Anopheles gambiae sl in Southern Benin

PubMed Central

Fongnikin, Augustin; Rowland, Mark; N’Guessan, Raphael

2017-01-01

Introduction There is an urgent need for new insecticides for indoor residual spraying (IRS) which can provide improved and prolonged control of malaria vectors that have developed resistance to existing insecticides. The neonicotinoid, clothianidin represents a class of chemistry new to public health. Clothianidin acts as an agonist on nicotinic acetyl choline receptors. IRS with a mixture of Clothianidin and another WHO approved insecticide such as deltamethrin could provide improved control of insecticide resistant malaria vector populations and serve as a tool for insecticide resistance management. Methods The efficacy and residual activity of a novel IRS mixture of deltamethrin and clothianidin was evaluated against wild pyrethroid resistant An. gambiae sl in experimental huts in Cove, Benin. Two application rates of the mixture were tested and comparison was made with clothianidin and deltamethrin applied alone. To assess the residual efficacy of the treatments on different local wall substrates, the inner walls of the experimental huts were covered with either cement, mud or plywood. Results Clothianidin demonstrated a clear delayed expression in mortality of wild pyrethroid resistant An. gambiae sl in the experimental huts which reached its full effect 120 hours after exposure. Overall mortality over the 12-month hut trial was 15% in the control hut and 24–29% in the deltamethrin-treated huts. The mixture of clothianidin 200mg/m2 and deltamethrin 25mg/m2 induced high overall hut mortality rates (87% on mud walls, 82% on cement walls and 61% on wooden walls) largely due to the clothianidin component and high hut exiting rates (67–76%) mostly due to the deltamethrin component. Mortality rates remained >80% for 8–9 months on mud and cement walls. The residual activity trend was confirmed by results from monthly in situ cone bioassays with laboratory susceptible An. gambiae Kisumu strain. Conclusion IRS campaigns with the mixture of clothianidin plus

Microbial Pre-exposure and Vectorial Competence of Anopheles Mosquitoes

PubMed Central

Dieme, Constentin; Rotureau, Brice; Mitri, Christian

2017-01-01

Anopheles female mosquitoes can transmit Plasmodium, the malaria parasite. During their aquatic life, wild Anopheles mosquito larvae are exposed to a huge diversity of microbes present in their breeding sites. Later, adult females often take successive blood meals that might also carry different micro-organisms, including parasites, bacteria, and viruses. Therefore, prior to Plasmodium ingestion, the mosquito biology could be modulated at different life stages by a suite of microbes present in larval breeding sites, as well as in the adult environment. In this article, we highlight several naturally relevant scenarios of Anopheles microbial pre-exposure that we assume might impact mosquito vectorial competence for the malaria parasite: (i) larval microbial exposures; (ii) protist co-infections; (iii) virus co-infections; and (iv) pathogenic bacteria co-infections. In addition, significant behavioral changes in African Anopheles vectors have been associated with increasing insecticide resistance. We discuss how these ethological modifications may also increase the repertoire of microbes to which mosquitoes could be exposed, and that might also influence their vectorial competence. Studying Plasmodium–Anopheles interactions in natural microbial environments would efficiently contribute to refining the transmission risks. PMID:29376030

How effective has tobacco tax increase been in the Gambia? A case study of tobacco control.

PubMed

Nargis, Nigar; Manneh, Yahya; Krubally, Bakary; Jobe, Baboucarr; Ouma, Ahmed E Ogwell; Tcha-Kondor, Noureiny; Blecher, Evan H

2016-08-26

The objective of the present study was to evaluate how effective tobacco tax increase has been in increasing price of tobacco products and reducing tobacco consumption in the Gambia. In addition, it tests the hypothesis that tobacco tax revenue grows while tobacco consumption decreases as a result of tax and price increase. The study is designed at the macroeconomic level to examine the import of tobacco products and revenue collected from tobacco taxation in a low-income setting. The participants of this study are the government officials employed in the Ministry of Finance and Economic Affairs (MoFEA), the Gambia and the Gambia Revenue Authority, who are in charge of planning and implementing the tobacco tax policy in the Gambia. The study includes 2 consecutive interventions in tobacco tax policy in the Gambia. The first intervention was moving the tax base for the uniform specific excise tax on cigarettes from weight to pack of cigarettes in 2013. The second intervention involved increasing the excise and the environmental tax on tobacco products in 2014. The primary outcome measures were the cost, insurance and freight value and the price of tobacco products. The secondary outcome measures included the import of tobacco products and tobacco tax revenue. In 2013-2014, the Gambia MoFEA raised the specific excise rate, which increased price, reduced consumption and generated significantly more government revenue from tobacco products. This is a clear evidence of the win-win outcome of raising tobacco tax. In addition, the Gambia has set the example of harmonising tax rates between tobacco products that reduces the substitution between tobacco products. The Gambia presents the best practice in tobacco taxation. There is need for documenting more country-specific evidence on the win-win outcome of raising tobacco tax. Published by the BMJ Publishing Group Limited. For permission to use (where not already granted under a licence) please go to http://www.bmj.com/company/products-services/rights-and-licensing/

Habitat suitability and ecological niche profile of major malaria vectors in Cameroon

PubMed Central

2009-01-01

Background Suitability of environmental conditions determines a species distribution in space and time. Understanding and modelling the ecological niche of mosquito disease vectors can, therefore, be a powerful predictor of the risk of exposure to the pathogens they transmit. In Africa, five anophelines are responsible for over 95% of total malaria transmission. However, detailed knowledge of the geographic distribution and ecological requirements of these species is to date still inadequate. Methods Indoor-resting mosquitoes were sampled from 386 villages covering the full range of ecological settings available in Cameroon, Central Africa. Using a predictive species distribution modeling approach based only on presence records, habitat suitability maps were constructed for the five major malaria vectors Anopheles gambiae, Anopheles funestus, Anopheles arabiensis, Anopheles nili and Anopheles moucheti. The influence of 17 climatic, topographic, and land use variables on mosquito geographic distribution was assessed by multivariate regression and ordination techniques. Results Twenty-four anopheline species were collected, of which 17 are known to transmit malaria in Africa. Ecological Niche Factor Analysis, Habitat Suitability modeling and Canonical Correspondence Analysis revealed marked differences among the five major malaria vector species, both in terms of ecological requirements and niche breadth. Eco-geographical variables (EGVs) related to human activity had the highest impact on habitat suitability for the five major malaria vectors, with areas of low population density being of marginal or unsuitable habitat quality. Sunlight exposure, rainfall, evapo-transpiration, relative humidity, and wind speed were among the most discriminative EGVs separating "forest" from "savanna" species. Conclusions The distribution of major malaria vectors in Cameroon is strongly affected by the impact of humans on the environment, with variables related to proximity to human

Efficacy of local neem extracts for sustainable malaria vector control in an African village

PubMed Central

Gianotti, Rebecca L; Bomblies, Arne; Dafalla, Mustafa; Issa-Arzika, Ibrahim; Duchemin, Jean-Bernard; Eltahir, Elfatih AB

2008-01-01

Background Larval control of malaria vectors has been historically successful in reducing malaria transmission, but largely fell out of favour with the introduction of synthetic insecticides and bed nets. However, an integrated approach to malaria control, including larval control methods, continues to be the best chance for success, in view of insecticide resistance, the behavioural adaptation of the vectors to changing environments and the difficulties of reaching the poorest populations most at risk,. Laboratory studies investigating the effects of neem seed (Azadirachta indica) extracts on Anopheles larvae have shown high rates of larval mortality and reductions in adult longevity, as well as low potential for resistance development. Methods This paper describes a method whereby seeds of the neem tree can be used to reduce adult Anopheles gambiae s.l. abundance in a way that is low cost and can be implemented by residents of rural villages in western Niger. The study was conducted in Banizoumbou village, western Niger. Neem seeds were collected from around the village. Dried seeds were ground into a coarse powder, which was then sprinkled onto known Anopheles larvae breeding habitats twice weekly during the rainy season 2007. Adult mosquitoes were captured on a weekly basis in the village and captures compared to those from 2005 and 2006 over the same period. Adult mosquitoes were also captured in a nearby village, Zindarou, as a control data set and compared to those from Banizoumbou. Results It was found that twice-weekly applications of the powder to known breeding habitats of Anopheles larvae in 2007 resulted in 49% fewer adult female Anopheles gambiae s.l. mosquitoes in Banizoumbou, compared with previous captures under similar environmental conditions and with similar habitat characteristics in 2005 and 2006. The productivity of the system in 2007 was found to be suppressed compared to the mean behaviour of 2005 and 2006 in Banizoumbou, whereas no change

Artemisinin-resistant Plasmodium falciparum clinical isolates can infect diverse mosquito vectors of Southeast Asia and Africa.

PubMed

St Laurent, Brandyce; Miller, Becky; Burton, Timothy A; Amaratunga, Chanaki; Men, Sary; Sovannaroth, Siv; Fay, Michael P; Miotto, Olivo; Gwadz, Robert W; Anderson, Jennifer M; Fairhurst, Rick M

2015-10-20

Artemisinin-resistant Plasmodium falciparum parasites are rapidly spreading in Southeast Asia, yet nothing is known about their transmission. This knowledge gap and the possibility that these parasites will spread to Africa endanger global efforts to eliminate malaria. Here we produce gametocytes from parasite clinical isolates that displayed artemisinin resistance in patients and in vitro, and use them to infect native and non-native mosquito vectors. We show that contemporary artemisinin-resistant isolates from Cambodia develop and produce sporozoites in two Southeast Asian vectors, Anopheles dirus and Anopheles minimus, and the major African vector, Anopheles coluzzii (formerly Anopheles gambiae M). The ability of artemisinin-resistant parasites to infect such highly diverse Anopheles species, combined with their higher gametocyte prevalence in patients, may explain the rapid expansion of these parasites in Cambodia and neighbouring countries, and further compromise efforts to prevent their global spread.

Screening Mosquito House Entry Points as a Potential Method for Integrated Control of Endophagic Filariasis, Arbovirus and Malaria Vectors

PubMed Central

Ogoma, Sheila B.; Lweitoijera, Dickson W.; Ngonyani, Hassan; Furer, Benjamin; Russell, Tanya L.; Mukabana, Wolfgang R.; Killeen, Gerry F.; Moore, Sarah J.

2010-01-01

Background Partial mosquito-proofing of houses with screens and ceilings has the potential to reduce indoor densities of malaria mosquitoes. We wish to measure whether it will also reduce indoor densities of vectors of neglected tropical diseases. Methodology The main house entry points preferred by anopheline and culicine vectors were determined through controlled experiments using specially designed experimental huts and village houses in Lupiro village, southern Tanzania. The benefit of screening different entry points (eaves, windows and doors) using PVC-coated fibre glass netting material in terms of reduced indoor densities of mosquitoes was evaluated compared to the control. Findings 23,027 mosquitoes were caught with CDC light traps; 77.9% (17,929) were Anopheles gambiae sensu lato, of which 66.2% were An. arabiensis and 33.8% An. gambiae sensu stricto. The remainder comprised 0.2% (50) An. funestus, 10.2% (2359) Culex spp. and 11.6% (2664) Mansonia spp. Screening eaves reduced densities of Anopheles gambiae s. l. (Relative ratio (RR)  = 0.91; 95% CI = 0.84, 0.98; P = 0.01); Mansonia africana (RR = 0.43; 95% CI = 0.26, 0.76; P<0.001) and Mansonia uniformis (RR = 0.37; 95% CI = 0.25, 0.56; P<0.001) but not Culex quinquefasciatus, Cx. univittatus or Cx. theileri. Numbers of these species were reduced by screening windows and doors but this was not significant. Significance This study confirms that across Africa, screening eaves protects households against important mosquito vectors of filariasis, Rift Valley Fever and O'Nyong nyong as well as malaria. While full house screening is required to exclude Culex species mosquitoes, screening of eaves alone or fitting ceilings has considerable potential for integrated control of other vectors of filariasis, arbovirus and malaria. PMID:20689815

ABC transporters are involved in defense against permethrin insecticide in the malaria vector Anopheles stephensi.

PubMed

Epis, Sara; Porretta, Daniele; Mastrantonio, Valentina; Comandatore, Francesco; Sassera, Davide; Rossi, Paolo; Cafarchia, Claudia; Otranto, Domenico; Favia, Guido; Genchi, Claudio; Bandi, Claudio; Urbanelli, Sandra

2014-07-29

Proteins from the ABC family (ATP-binding cassette) represent the largest known group of efflux pumps, responsible for transporting specific molecules across lipid membranes in both prokaryotic and eukaryotic organisms. In arthropods they have been shown to play a role in insecticide defense/resistance. The presence of ABC transporters and their possible association with insecticide transport have not yet been investigated in the mosquito Anopheles stephensi, the major vector of human malaria in the Middle East and South Asian regions. Here we investigated the presence and role of ABCs in transport of permethrin insecticide in a susceptible strain of this mosquito species. To identify ABC transporter genes we obtained a transcriptome from untreated larvae of An. stephensi and then compared it with the annotated transcriptome of Anopheles gambiae. To analyse the association between ABC transporters and permethrin we conducted bioassays with permethrin alone and in combination with an ABC inhibitor, and then we investigated expression profiles of the identified genes in larvae exposed to permethrin. Bioassays showed an increased mortality of mosquitoes when permethrin was used in combination with the ABC-transporter inhibitor. Genes for ABC transporters were detected in the transcriptome, and five were selected (AnstABCB2, AnstABCB3, AnstABCB4, AnstABCmember6 and AnstABCG4). An increased expression in one of them (AnstABCG4) was observed in larvae exposed to the LD50 dose of permethrin. Contrary to what was found in other insect species, no up-regulation was observed in the AnstABCB genes. Our results show for the first time the involvement of ABC transporters in larval defense against permethrin in An. stephensi and, more in general, confirm the role of ABC transporters in insecticide defense. The differences observed with previous studies highlight the need of further research as, despite the growing number of studies on ABC transporters in insects, the

Genomic, Physiologic, and Symbiotic Characterization of Serratia marcescens Strains Isolated from the Mosquito Anopheles stephensi.

PubMed

Chen, Shicheng; Blom, Jochen; Walker, Edward D

2017-01-01

Strains of Serratia marcescens , originally isolated from the gut lumen of adult female Anopheles stephensi mosquitoes, established persistent infection at high rates in adult A. stephensi whether fed to larvae or in the sugar meal to adults. By contrast, the congener S. fonticola originating from Aedes triseriatus had lower infection in A. stephensi , suggesting co-adaptation of Serratia strains in different species of host mosquitoes. Coinfection at high infection rate in adult A. stephensi resulted after feeding S. marcescens and Elizabethkingia anophelis in the sugar meal, but when fed together to larvae, infection rates with E. anophelis were much higher than were S. marcescens in adult A. stephensi , suggesting a suppression effect of coinfection across life stages. A primary isolate of S. marcescens was resistant to all tested antibiotics, showed high survival in the mosquito gut, and produced alpha-hemolysins which contributed to lysis of erythrocytes ingested with the blood meal. Genomes of two primary isolates from A. stephensi , designated S. marcescens ano1 and ano2, were sequenced and compared to other Serratia symbionts associated with insects, nematodes and plants. Serratia marcescens ano1 and ano2 had predicted virulence factors possibly involved in attacking parasites and/or causing opportunistic infection in mosquito hosts. S. marcescens ano1 and ano2 possessed multiple mechanisms for antagonism against other microorganisms, including production of bacteriocins and multi-antibiotic resistance determinants. These genes contributing to potential anti-malaria activity including serralysins, hemolysins and chitinases are only found in some Serratia species. It is interesting that genome sequences in S. marcescens ano1 and ano2 are distinctly different from those in Serratia sp. Ag1 and Ag2 which were isolated from Anopheles gambiae . Compared to Serratia sp. Ag1 and Ag2, S. marcescens ano1 and ano2 have more rRNAs and many important genes involved in

Genomic, Physiologic, and Symbiotic Characterization of Serratia marcescens Strains Isolated from the Mosquito Anopheles stephensi

PubMed Central

Chen, Shicheng; Blom, Jochen; Walker, Edward D.

2017-01-01

Strains of Serratia marcescens, originally isolated from the gut lumen of adult female Anopheles stephensi mosquitoes, established persistent infection at high rates in adult A. stephensi whether fed to larvae or in the sugar meal to adults. By contrast, the congener S. fonticola originating from Aedes triseriatus had lower infection in A. stephensi, suggesting co-adaptation of Serratia strains in different species of host mosquitoes. Coinfection at high infection rate in adult A. stephensi resulted after feeding S. marcescens and Elizabethkingia anophelis in the sugar meal, but when fed together to larvae, infection rates with E. anophelis were much higher than were S. marcescens in adult A. stephensi, suggesting a suppression effect of coinfection across life stages. A primary isolate of S. marcescens was resistant to all tested antibiotics, showed high survival in the mosquito gut, and produced alpha-hemolysins which contributed to lysis of erythrocytes ingested with the blood meal. Genomes of two primary isolates from A. stephensi, designated S. marcescens ano1 and ano2, were sequenced and compared to other Serratia symbionts associated with insects, nematodes and plants. Serratia marcescens ano1 and ano2 had predicted virulence factors possibly involved in attacking parasites and/or causing opportunistic infection in mosquito hosts. S. marcescens ano1 and ano2 possessed multiple mechanisms for antagonism against other microorganisms, including production of bacteriocins and multi-antibiotic resistance determinants. These genes contributing to potential anti-malaria activity including serralysins, hemolysins and chitinases are only found in some Serratia species. It is interesting that genome sequences in S. marcescens ano1 and ano2 are distinctly different from those in Serratia sp. Ag1 and Ag2 which were isolated from Anopheles gambiae. Compared to Serratia sp. Ag1 and Ag2, S. marcescens ano1 and ano2 have more rRNAs and many important genes involved in

Molecular epidemiology of respiratory syncytial virus in The Gambia.

PubMed Central

Cane, P. A.; Weber, M.; Sanneh, M.; Dackour, R.; Pringle, C. R.; Whittle, H.

1999-01-01

Respiratory syncytial virus (RSV) infection in The Gambia occurs seasonally in association with the rainy season. This study examined the genetic variability of RSV isolates from four consecutive epidemics from 1993-6. Each epidemic was made up of a number of variants which were replaced in subsequent epidemics. Analysis of attachment (G) protein gene sequences showed that isolates were closely related to those observed in the rest of the world. However, many isolates from 1993 and 1994 were unlike other isolates observed in the developed world during this period and were more similar to isolates from 1984 in Europe. In addition, the most commonly observed genotype in the UK in the 1990s was not detected in The Gambia during this period. PMID:10098799

76 FR 61133 - Waiver of Restriction on Assistance to the Central Government of The Gambia

Federal Register 2010, 2011, 2012, 2013, 2014

2011-10-03

... DEPARTMENT OF STATE [Public Notice: 7625] Waiver of Restriction on Assistance to the Central Government of The Gambia Pursuant to Section 7086(c)(2) of the Department of State, Foreign Operations, and... The Gambia and I hereby waive such restriction. This determination shall be reported to the Congress...

Screening of pesticide residues in soil and water samples from agricultural settings

PubMed Central

Akogbéto, Martin C; Djouaka, Rousseau F; Kindé-Gazard, Dorothée A

2006-01-01

Background The role of agricultural practices in the selection of insecticide resistance in malaria vectors has so far been hypothesized without clear evidence. Many mosquito species, Anopheles gambiae in particular, lay their eggs in breeding sites located around agricultural settings. There is a probability that, as a result of farming activities, insecticide residues may be found in soil and water, where they exercise a selection pressure on the larval stage of various populations of mosquitoes. To confirm this hypothesis, a study was conducted in the Republic of Benin to assess the environmental hazards which can be generated from massive use of pesticides in agricultural settings. Methods Lacking an HPLC machine for direct quantification of insecticide residues in samples, this investigation was performed using indirect bioassays focussed on the study of factors inhibiting the normal growth of mosquito larvae in breeding sites. The speed of development was monitored as well as the yield of rearing An. gambiae larvae in breeding sites reconstituted with water and soil samples collected in agricultural areas known to be under pesticide pressure. Two strains of An. gambiae were used in this indirect bioassay: the pyrethroid-susceptible Kisumu strain and the resistant Ladji strain. The key approach in this methodology is based on comparison of the growth of larvae in test and in control breeding sites, the test samples having been collected from two vegetable farms. Results Results obtained clearly show the presence of inhibiting factors on test samples. A normal growth of larvae was observed in control samples. In breeding sites simulated by using a few grams of soil samples from the two vegetable farms under constant insecticide treatments (test samples), a poor hatching rate of Anopheles eggs coupled with a retarded growth of larvae and a low yield of adult mosquitoes from hatched eggs, was noticed. Conclusion Toxic factors inhibiting the hatching of anopheles

78 FR 76699 - Waiver of Restriction on Assistance to the Central Government of The Gambia

Federal Register 2010, 2011, 2012, 2013, 2014

2013-12-18

... DEPARTMENT OF STATE [Public Notice 8555] Waiver of Restriction on Assistance to the Central Government of The Gambia Pursuant to Section 7031(b)(3) of the Department of State, Foreign Operations, and... similar provisions of law in prior year Acts with respect to The Gambia and I hereby waive this...

Fluoride availability from natural resources in The Gambia--implications for oral health care.

PubMed

Jordan, Rainer A; Markovic, Ljubisa; Gaengler, Peter

2008-10-01

Changing food patterns in combination with ineffective oral hygiene measures and insufficient bioavailability of fluoride from drinking water and other sources seem to impair the dental health status in developing countries, especially in the younger population. Therefore, preventive programmes in controlling dental caries progression should be based on local conditions. For mapping the drinking water fluoride content throughout The Gambia, samples of water from rural community wells, public water taps, commercial mineral water, and from the Gambia-River were measured. Additionally, fluoride concentrations of locally extracted table salt and green tea were determined. Showed the need for supplementary fluoride intake, because natural dietary fluoride availability is very low. Age-related recommendations for oral health care and for additional fluoride bioavailability are given, taking into account local socio-economic conditions in the Republic of The Gambia and similar developing countries.

Environmentally friendly tool to control mosquito populations without risk of insecticide resistance: the Lehmann’s funnel entry trap

PubMed Central

2013-01-01

Background Current malaria control strategies have cut down the malaria burden in many endemic areas, however the emergence and rapid spread of insecticide and drug resistance undermine the success of these efforts. There is growing concern that malaria eradication will not be achieved without the introduction of novel control tools. One approach that has been developed in the last few years is based on house screening to reduce indoor mosquito vector densities and consequently decrease malaria transmission. Here screening and trapping were combined in one tool to control mosquito populations. The trap does not require an insecticide or even an attractant, yet it effectively collects incoming resistant and susceptible mosquitoes and kills them. Results Performance of the funnel entry trap was tested in low and high malaria vector density areas. An overall reduction of 70 to 80% of mosquito density was seen in both. Species and molecular forms of Anopheles gambiae identification indicated no variation in the number of Anopheles arabiensis and the molecular forms of An. gambiae between houses and traps. Mosquitoes collected in the traps and in houses were highly resistant to pyrethroids (0.9 kdr-based mechanism). Conclusion There is a global consensus that new intervention tools are needed to cross the last miles in malaria elimination/eradication. The funnel entry trap showed excellent promise in suppressing mosquito densities even in area of high insecticide resistance. It requires no chemicals and is self-operated. PMID:23758904

Chronic non-communicable disease as a new epidemic in Africa: focus on The Gambia

PubMed Central

Omoleke, Semeeh A

2013-01-01

Introduction Recent epidemiological data suggest increasing burden of NCDs in many African countries but these diseases have not been given adequate attention due to the overwhelming burden of infectious diseases. There are no recent reports or studies on NCDs or related issues in The Gambia, consequently, this report intends to stimulate further epidemiological studies and also policy initiatives to forestall an epidemic. Methods Routine data on morbidity (in and out-patients), hospitalisation and mortality due to NCDs from health facilities in The Gambia between 2008 and 2011 were used. Other relevant data from multiple sources were also used. Results There is an increasing trend in the morbidity, hospitalisation and mortality due to NCDs in the Gambia between 2008 and 2011; 19.8%, 9.9% and 23.4% increments respectively. There is evidence of gender differences in these variables; more males suffer higher mortality from NCDs than females (p < 0.001). Furthermore, there is dearth of highly skilled health workforce as well as poor health infrastructures in The Gambia. Conclusion NCDs are becoming a public health challenge and the capacity to respond to NCDs in most African countries, particularly, The Gambia is very weak. There is need for a population-based study to accurately quantify the burden and their risk factors as a first step towards policy formulation and effective implementation. Furthermore, there is dire need for increased investments on health workforce as well as medical products and technologies towards addressing the consequences of this emerging epidemic. PMID:23646223

Genome-wide and expression-profiling analyses suggest the main cytochrome P450 genes related to pyrethroid resistance in the malaria vector, Anopheles sinensis (Diptera Culicidae).

PubMed

Yan, Zheng-Wen; He, Zheng-Bo; Yan, Zhen-Tian; Si, Feng-Ling; Zhou, Yong; Chen, Bin

2018-02-02

Anopheles sinensis is one of the major malaria vectors. However, pyrethroid resistance in An. sinensis is threatening malaria control. Cytochrome P450-mediated detoxification is an important pyrethroid resistance mechanism that has been unexplored in An. sinensis. In this study, we performed a comprehensive analysis of the An. sinensis P450 gene superfamily with special attention to their role in pyrethroid resistance using bioinformatics and molecular approaches. Our data revealed the presence of 112 individual P450 genes in An. sinensis, which were classified into four major clans (mitochondrial, CYP2, CYP3 and CYP4), 18 families and 50 subfamilies. Sixty-seven genes formed nine gene clusters, and genes within the same cluster and the same gene family had a similar gene structure. Phylogenetic analysis showed that most of An. sinensis P450s (82/112) had very close 1: 1 orthology with Anopheles gambiae P450s. Five genes (AsCYP6Z2, AsCYP6P3v1, AsCYP6P3v2, AsCYP9J5 and AsCYP306A1) were significantly upregulated in three pyrethroid-resistant populations in both RNA-seq and RT-qPCR analyses, suggesting that they could be the most important P450 genes involved in pyrethroid resistance in An. sinensis. Our study provides insight on the diversity of An. sinensis P450 superfamily and basis for further elucidating pyrethroid resistance mechanism in this mosquito species. © 2018 Society of Chemical Industry. © 2018 Society of Chemical Industry.

Artemisinin-resistant Plasmodium falciparum clinical isolates can infect diverse mosquito vectors of Southeast Asia and Africa

PubMed Central

St. Laurent, Brandyce; Miller, Becky; Burton, Timothy A.; Amaratunga, Chanaki; Men, Sary; Sovannaroth, Siv; Fay, Michael P.; Miotto, Olivo; Gwadz, Robert W.; Anderson, Jennifer M.; Fairhurst, Rick M.

2015-01-01

Artemisinin-resistant Plasmodium falciparum parasites are rapidly spreading in Southeast Asia, yet nothing is known about their transmission. This knowledge gap and the possibility that these parasites will spread to Africa endanger global efforts to eliminate malaria. Here we produce gametocytes from parasite clinical isolates that displayed artemisinin resistance in patients and in vitro, and use them to infect native and non-native mosquito vectors. We show that contemporary artemisinin-resistant isolates from Cambodia develop and produce sporozoites in two Southeast Asian vectors, Anopheles dirus and Anopheles minimus, and the major African vector, Anopheles coluzzii (formerly Anopheles gambiae M). The ability of artemisinin-resistant parasites to infect such highly diverse Anopheles species, combined with their higher gametocyte prevalence in patients, may explain the rapid expansion of these parasites in Cambodia and neighbouring countries, and further compromise efforts to prevent their global spread. PMID:26485448

Diversity of Anopheles mosquitoes in Binh Phuoc and Dak Nong Provinces of Vietnam and their relation to disease

PubMed Central

2014-01-01

Background Human malaria is still a burden in Dak Nong and Binh Phuoc Provinces in south-central Vietnam that border Cambodia. Several Anopheles species that transmit human malarial Plasmodium may also transmit Wuchereria bancrofti, the nematode that causes Bancroftian lymphatic filariasis. The objective of this study was to investigate the role of Anopheles species in the transmission of these two pathogens in the two highly malaria endemic provinces of Vietnam. Methods Anopheles mosquitoes were collected in Dak Nong and Binh Phuoc Provinces in November and December of 2010 and 2011. Human landing catches, paired collections on human and buffalo, and resting captures were made with mouth aspirators. Collections were also made with light traps. Morphological and PCR-based methods were used to identify the species. Real-time PCR was used to detect Plasmodium species and W. bancrofti in individual mosquitoes. Results Twenty-four Anopheles species were identified among 797 captured mosquitoes. Anopheles dirus was found in both provinces and was the predominant species in Binh Phuoc Province; An. maculatus was the most prevalent species in Dak Nong Province. Anopheles minimus was collected only in Binh Phuoc Province. Some specimens of An. minimus and An. pampanai were misidentified based on morphology. Four specimens of An. scanloni were identified, and this is the first report of this species of the Dirus Complex in Vietnam. Two females, one An. dirus and one An. pampanai, collected in Binh Phuoc Province were infected with P. vivax, for an overall infection rate of 0.41% (2/486): 0.28% for An. dirus (1/361) and 20% for An. pampanai (1/5). No mosquitoes were found to be infected with P. falciparum, P. knowlesi or W. bancrofti in either province. Conclusion A diversity of Anopheles species occurs in Dak Nong and Binh Phuoc Provinces of Vietnam, several of which are considered to be actual and potential vectors of malarial protozoa and microfilariae. It is highly

The resting sites and blood-meal sources of Anopheles minimus in Taiwan.

PubMed

Chang, Mei-Chun; Teng, Hwa-Jen; Chen, Chen-Fu; Chen, Yung-Chen; Jeng, Chian-Ren

2008-06-09

The WHO declared Taiwan free from malaria in 1965, but in 2003 the reporting of two introduced cases in a rural area suggested a possible local transmission of this disease. Therefore, understanding the resting sites and the blood sources of Anopheles minimus is crucial in order to provide information for implementing vector control strategies. During a two-year survey, mosquitoes were collected in houses and their surrounding areas and at the bank of larval habitats by backpack aspirators in 17 villages in rural areas of southern and eastern Taiwan for 1 hr. On the same day, blacklight traps were hung downward overnight. Blood-fed mosquito samples were analysed by PCR. Of the 195 total households surveyed by backpack aspirators, no Anopheles adults were collected inside the houses, while a single Anopheles minimus and a single Anopheles maculatus were collected outside of the houses. On the same day, 23 An. minimus, two An. maculatus, two Anopheles ludlowae, two Anopheles sinensis, and one Anopheles tessellatus were collected along the bank of larval habitats. In blacklight traps hung outside of the houses in the villages, 69 An. minimus, 62 An. ludlowae, 31 An. sinensis, and 19 An. maculatus were collected. In larval habitats, 98 An. ludlowae, 64 An. minimus, 49 An. sinensis, and 14 An. maculatus were collected. Of a total of 10 blood-fed samples, An. minimus fed on four animals including bovine (60%), dogs (20%), pig (10%), and non-chicken avian (10%). Anopheles minimus, an opportunist feeder in Taiwan, was not collected inside the houses, but was found outside of the houses in villages and surrounding larval habitats. Therefore, an outdoor transmission of malaria is likely to occur and, thus, the bed nets, which are favoured for controlling the late biting of An. minimus, should be a very efficient and effective method for those local residents who sleep outdoors. Additionally, space spray of insecticides for Anopheles at night, as well as residual spray inside

Reconstructing the flight kinematics of swarming and mating behavior in wild mosquitoes

USDA-ARS?s Scientific Manuscript database

We describe a tracking system for reconstructing three-dimensional tracks of individual mosquitoes in wild swarms and present the results of validating the system by filming swarms and mating events of the malaria mosquito Anopheles gambiae in Mali. The tracking system is designed to address noisy, ...

Genome-wide identification, characterization and classification of ionotropic glutamate receptor genes (iGluRs) in the malaria vector Anopheles sinensis (Diptera: Culicidae).

PubMed

Wang, Ting-Ting; Si, Feng-Ling; He, Zheng-Bo; Chen, Bin

2018-01-15

Ionotropic glutamate receptors (iGluRs) are conserved ligand-gated ion channel receptors, and ionotropic receptors (IRs) were revealed as a new family of iGluRs. Their subdivision was unsettled, and their characteristics are little known. Anopheles sinensis is a major malaria vector in eastern Asia, and its genome was recently well sequenced and annotated. We identified iGluR genes in the An. sinensis genome, analyzed their characteristics including gene structure, genome distribution, domains and specific sites by bioinformatic methods, and deduced phylogenetic relationships of all iGluRs in An. sinensis, Anopheles gambiae and Drosophila melanogaster. Based on the characteristics and phylogenetics, we generated the classification of iGluRs, and comparatively analyzed the intron number and selective pressure of three iGluRs subdivisions, iGluR group, Antenna IR and Divergent IR subfamily. A total of 56 iGluR genes were identified and named in the whole-genome of An. sinensis. These genes were located on 18 scaffolds, and 31 of them (29 being IRs) are distributed into 10 clusters that are suggested to form mainly from recent gene duplication. These iGluRs can be divided into four groups: NMDA, non-NMDA, Antenna IR and Divergent IR based on feature comparison and phylogenetic analysis. IR8a and IR25a were suggested to be monophyletic, named as Putative in the study, and moved from the Antenna subfamily in the IR family to the non-NMDA group as a sister of traditional non-NMDA. The generated iGluRs of genes (including NMDA and regenerated non-NMDA) are relatively conserved, and have a more complicated gene structure, smaller ω values and some specific functional sites. The iGluR genes in An. sinensis, An. gambiae and D. melanogaster have amino-terminal domain (ATD), ligand binding domain (LBD) and Lig_Chan domains, except for IR8a that only has the LBD and Lig_Chan domains. However, the new concept IR family of genes (including regenerated Antenna IR, and Divergent

Confirmation of Anopheles peditaeniatus and Anopheles sundaicus as Malaria Vectors (Diptera: Culicidae) in Sungai Nyamuk Village, Sebatik Island North Kalimantan, Indonesia Using an Enzyme-Linked Immunosorbent Assay.

PubMed

Sugiarto; Kesumawati Hadi, Upik; Soviana, Susi; Hakim, Lukman

2016-11-01

Anopheles mosquitoes may be incriminated as malaria vectors by observing sporozoites in their salivary glands and by testing heads or thoraces by an enzyme-linked immunosorbent assay (ELISA) to detect Plasmodium species-specific circumsporozoite proteins (CSP). This study tested Anopheles collected in Sungai Nyamuk Village for the presence of both Plasmodium falciparum and Plasmodium vivax CSP. The Anopheles spp. were collected by human landing collection indoors and outdoors and by indoor and outdoor resting catches in Sungai Nyamuk Village, Nunukan District, North Kalimantan Province from August 2010 to January 2012. Overall, 5,100 Anopheles spp. comprising 11 species were collected and 2,259 adult parous females were tested by ELISA. Of these, only one Anopheles peditaeniatus Leicester (3.8%, n = 26) and one Anopheles sundaicus sensu lato (0.6%, n = 157) that originated from outdoor biting catches tested positive for P. falciparum CSP. The remaining females from indoor biting, outdoor resting, and indoor resting catches were negative for P. falciparum and P. vivax proteins. Confirmation of these vectors biting outdoors indicated that P. falciparum transmission may be occurring outside of houses by An. peditaeniatus and An. sundaicus. © The Authors 2016. Published by Oxford University Press on behalf of Entomological Society of America. All rights reserved. For Permissions, please email: journals.permissions@oup.com.

Thermal behaviour of Anopheles stephensi in response to infection with malaria and fungal entomopathogens

PubMed Central

Blanford, Simon; Read, Andrew F; Thomas, Matthew B

2009-01-01

Background Temperature is a critical determinant of the development of malaria parasites in mosquitoes, and hence the geographic distribution of malaria risk, but little is known about the thermal preferences of Anopheles. A number of other insects modify their thermal behaviour in response to infection. These alterations can be beneficial for the insect or for the infectious agent. Given current interest in developing fungal biopesticides for control of mosquitoes, Anopheles stephensi were examined to test whether mosquitoes showed thermally-mediated behaviour in response to infection with fungal entomopathogens and the rodent malaria, Plasmodium yoelii. Methods Over two experiments, groups of An. stephensi were infected with one of three entomopathogenic fungi, and/or P. yoelii. Infected and uninfected mosquitoes were released on to a thermal gradient (14 – 38°C) for "snapshot" assessments of thermal preference during the first five days post-infection. Mosquito survival was monitored for eight days and, where appropriate, oocyst prevalence and intensity was assessed. Results and conclusion Both infected and uninfected An. stephensi showed a non-random distribution on the gradient, indicating some capacity to behaviourally thermoregulate. However, chosen resting temperatures were not altered by any of the infections. There is thus no evidence that thermally-mediated behaviours play a role in determining malaria prevalence or that they will influence the performance of fungal biopesticides against adult Anopheles. PMID:19379519

Molecular Profiling of Phagocytic Immune Cells in Anopheles gambiae Reveals Integral Roles for Hemocytes in Mosquito Innate Immunity*

PubMed Central

Smith, Ryan C.; King, Jonas G.; Tao, Dingyin; Zeleznik, Oana A.; Brando, Clara; Thallinger, Gerhard G.; Dinglasan, Rhoel R.

2016-01-01

The innate immune response is highly conserved across all eukaryotes and has been studied in great detail in several model organisms. Hemocytes, the primary immune cell population in mosquitoes, are important components of the mosquito innate immune response, yet critical aspects of their biology have remained uncharacterized. Using a novel method of enrichment, we isolated phagocytic granulocytes and quantified their proteomes by mass spectrometry. The data demonstrate that phagocytosis, blood-feeding, and Plasmodium falciparum infection promote dramatic shifts in the proteomic profiles of An. gambiae granulocyte populations. Of interest, large numbers of immune proteins were induced in response to blood feeding alone, suggesting that granulocytes have an integral role in priming the mosquito immune system for pathogen challenge. In addition, we identify several granulocyte proteins with putative roles as membrane receptors, cell signaling, or immune components that when silenced, have either positive or negative effects on malaria parasite survival. Integrating existing hemocyte transcriptional profiles, we also compare differences in hemocyte transcript and protein expression to provide new insight into hemocyte gene regulation and discuss the potential that post-transcriptional regulation may be an important component of hemocyte gene expression. These data represent a significant advancement in mosquito hemocyte biology, providing the first comprehensive proteomic profiling of mosquito phagocytic granulocytes during homeostasis blood-feeding, and pathogen challenge. Together, these findings extend current knowledge to further illustrate the importance of hemocytes in shaping mosquito innate immunity and their principal role in defining malaria parasite survival in the mosquito host. PMID:27624304

Identification of novel cytochrome P450s in the Acari

USDA-ARS?s Scientific Manuscript database

Cytochrome P450s are the major phase I drug metabolising enzymes found in most organisms, including arthropods. Much of the work within the area of xenobiotic metabolism in this group of animals has centered around mosquito species, e.g. Anopheles gambiae and Culex quinquefasciatus, due to their rol...

Habitat segregation and ecological character displacement in cryptic African malaria mosquitoes

PubMed Central

Tene Fossog, Billy; Ayala, Diego; Acevedo, Pelayo; Kengne, Pierre; Ngomo Abeso Mebuy, Ignacio; Makanga, Boris; Magnus, Julie; Awono-Ambene, Parfait; Njiokou, Flobert; Pombi, Marco; Antonio-Nkondjio, Christophe; Paupy, Christophe; Besansky, Nora J; Costantini, Carlo

2015-01-01

Understanding how divergent selection generates adaptive phenotypic and population diversification provides a mechanistic explanation of speciation in recently separated species pairs. Towards this goal, we sought ecological gradients of divergence between the cryptic malaria vectors Anopheles coluzzii and An. gambiae and then looked for a physiological trait that may underlie such divergence. Using a large set of occurrence records and eco-geographic information, we built a distribution model to predict the predominance of the two species across their range of sympatry. Our model predicts two novel gradients along which the species segregate: distance from the coastline and altitude. Anopheles coluzzii showed a ‘bimodal’ distribution, predominating in xeric West African savannas and along the western coastal fringe of Africa. To test whether differences in salinity tolerance underlie this habitat segregation, we assessed the acute dose–mortality response to salinity of thirty-two larval populations from Central Africa. In agreement with its coastal predominance, Anopheles coluzzii was overall more tolerant than An. gambiae. Salinity tolerance of both species, however, converged in urban localities, presumably reflecting an adaptive response to osmotic stress from anthropogenic pollutants. When comparing degree of tolerance in conjunction with levels of syntopy, we found evidence of character displacement in this trait. PMID:25926878

Pesticide handling and exposures among cotton farmers in the gambia.

PubMed

Kuye, Rex A; Donham, Kelley J; Marquez, Shannon P; Sanderson, Wayne T; Fuortes, Laurence J; Rautiainen, Risto H; Jones, Martin L; Culp, Kennith R

2007-01-01

There are substantial health hazards to farmers and the environment associated with pesticide use in developing countries. Based on observations by the authors and previous reports, most previous studies in Africa are descriptive in nature. The aim of this study was to investigate how cotton farmers are exposed to pesticides in The Gambia and quantify their pesticide exposures and provide information for the formulation of a policy on pesticide safety for the country. A representative sample of 20 cotton farmers in the Central and Upper River Divisions (CRD and URD) of The Gambia were surveyed by questionnaires. Dermal pesticide exposures among a subset of 10 farmer/pesticide applicators were assessed by dermal patch samples, observation, and postapplication questionnaires. The study revealed that a toxic organochlorine insecticide, Callisulfan (endosulfan), is frequently sprayed on cotton plants by the farmers. The farmers wore no protective equipment and were inadequately dressed for work with this pesticide. Laboratory analysis of the mixed formulation showed a wide range in the concentration of the pesticide solution among the farmer/pesticide applicators and dermal patch samples showed very high residues of endosulfan analytes on their body surfaces. A low level of awareness of pesticide toxicity prevails amonsg cotton farmers in The Gambia. There is a less than adequate control of pesticides and other hazardous agrichemicals in the country.

Agriculture and the promotion of insect pests: rice cultivation in river floodplains and malaria vectors in The Gambia.

PubMed

Jarju, Lamin B S; Fillinger, Ulrike; Green, Clare; Louca, Vasilis; Majambere, Silas; Lindsay, Steven W

2009-07-27

Anthropogenic modification of natural habitats can create conditions in which pest species associated with humans can thrive. In order to mitigate for these changes, it is necessary to determine which aspects of human management are associated with the promotion of those pests. Anopheles gambiae, the main Africa malaria vector, often breeds in rice fields. Here the impact of the ancient practice of 'swamp rice' cultivation, on the floodplains of the Gambia River, on the production of anopheline mosquitoes was investigated. Routine surveys were carried out along 500 m transects crossing rice fields from the landward edge of the floodplains to the river during the 2006 rainy season. Aquatic invertebrates were sampled using area samplers and emergence traps and fish sampled using nets. Semi-field experiments were used to investigate whether nutrients used for swamp rice cultivation affected mosquito larval abundance. At the beginning of the rainy season rice is grown on the landward edge of the floodplain; the first area to flood with fresh water and one rich in cattle dung. Later, rice plants are transplanted close to the river, the last area to dry out on the floodplain. Nearly all larval and adult stages of malaria vectors were collected 0-100 m from the landward edge of the floodplains, where immature rice plants were grown. These paddies contained stagnant freshwater with high quantities of cattle faeces. Semi-field studies demonstrated that cattle faeces nearly doubled the number of anopheline larvae compared with untreated water. Swamp rice cultivation creates ideal breeding sites for malaria vectors. However, only those close to the landward edge harboured vectors. These sites were productive since they were large areas of standing freshwater, rich in nutrients, protected from fish, and situated close to human habitation, where egg-laying mosquitoes from the villages had short distances to fly. The traditional practice of 'swamp rice' cultivation uses

Comparative evaluation of the Ifakara tent trap-B, the standardized resting boxes and the human landing catch for sampling malaria vectors and other mosquitoes in urban Dar es Salaam, Tanzania

PubMed Central

Sikulu, Maggy; Govella, Nicodem J; Ogoma, Sheila B; Mpangile, John; Kambi, Said H; Kannady, Khadija; Chaki, Prosper C; Mukabana, Wolfgang R; Killeen, Gerry F

2009-01-01

Background Frequent, sensitive and accurate sampling of Anopheles mosquitoes is a prerequisite for effective management of malaria vector control programmes. The most reliable existing means to measure mosquito density is the human landing catch (HLC). However, the HLC technique raises major ethical concerns because of the necessity to expose humans to vectors of malaria and a variety of other pathogens. Furthermore, it is a very arduous undertaking that requires intense supervision, which is severely limiting in terms of affordability and sustainability. Methods A community-based, mosquito sampling protocol, using the Ifakara tent trap-B (ITT-B) and standardized resting boxes (SRB), was developed and evaluated in terms of the number and sample composition of mosquitoes caught by each, compared to rigorously controlled HLC. Mosquitoes were collected once and three times every week by the HLC and the alternative methods, respectively, in the same time and location. Results Overall, the three traps caught 44,848 mosquitoes. The ITT-B, HLC and SRB caught 168, 143 and 46 Anopheles gambiae s.l. as well as 26,315, 13,258 and 4,791 Culex species respectively. The ITT-B was three- and five-times cheaper than the HLC per mosquito caught for An. gambiae and Cx. Species, respectively. Significant correlations between the numbers caught by HLC and ITT-B were observed for both An. gambiae s.l. (P < 0.001) and Cx. species (P = 0.003). Correlation between the catches with HLC and SRB were observed for Cx. species (P < 0.001) but not An. gambiae s.l. (P = 0.195), presumably because of the low density of the latter. Neither ITT-B nor SRB exhibited any obvious density dependence for sampling the two species. Conclusion SRBs exhibited poor sensitivity for both mosquito taxa and are not recommended in this setting. However, this protocol is affordable and effective for routine use of the ITT-B under programmatic conditions. Nevertheless, it is recommended that the trap and the

Existence of the rdl mutant alleles among the anopheles malaria vector in Indonesia.

PubMed

Asih, Puji Bs; Syahrani, Lepa; Rozi, Ismail Ep; Pratama, Nandha R; Marantina, Sylvia S; Arsyad, Dian S; Mangunwardoyo, Wibowo; Hawley, William; Laihad, Ferdinand; Shinta; Sukowati, Supratman; Lobo, Neil F; Syafruddin, Din

2012-02-25

The gamma-aminobutyric acid (GABA) receptor-chloride channel complex is known to be the target site of dieldrin, a cyclodiene insecticide. GABA-receptors, with a naturally occurring amino acid substitution, A302S/G in the putative ion-channel lining region, confer resistance to cyclodiene insecticides that includes aldrin, chlordane, dieldrin, heptachlor, endrin and endosulphan. A total of 154 mosquito samples from 10 provinces of malaria-endemic areas across Indonesia (Aceh, North Sumatra, Bangka Belitung, Lampung, Central Java, East Nusa Tenggara, West Nusa Tenggara, West Sulawesi, Molucca and North Molucca) were obtained and identified by species, using morphological characteristic. The DNA was individually extracted using chelex-ion exchanger and the DNA obtained was used for analyses using sequencing method. Molecular analysis indicated 11% of the total 154 Anopheles samples examined, carried Rdl mutant alleles. All of the alleles were found in homozygous form. Rdl 302S allele was observed in Anopheles vagus (from Central Java, Lampung, and West Nusa Tenggara), Anopheles aconitus (from Central Java), Anopheles barbirostris (from Central Java and Lampung), Anopheles sundaicus (from North Sumatra and Lampung), Anopheles nigerrimus (from North Sumatra), whereas the 302 G allele was only found in Anopheles farauti from Molucca. The existence of the Rdl mutant allele indicates that, either insecticide pressure on the Anopheles population in these areas might still be ongoing (though not directly associated with the malaria control programme) or that the mutant form of the Rdl allele is relatively stable in the absence of insecticide. Nonetheless, the finding suggests that integrated pest management is warranted in malaria-endemic areas where insecticides are widely used for other purposes.

Girls' Familial Responsibilities and Schooling in The Gambia

ERIC Educational Resources Information Center

Njie, Haddy; Manion, Caroline; Badjie, Musukuta

2015-01-01

Like many countries in the developing world gender inequity remains a staggering problem in The Gambia, particularly at the secondary school level. In this study, we focus on the relationship between girls' education and heavy domestic workloads, herein referred to as girls' familial responsibilities. We explore this topic in relation not only to…

Anopheles Vectors in Mainland China While Approaching Malaria Elimination.

PubMed

Zhang, Shaosen; Guo, Shaohua; Feng, Xinyu; Afelt, Aneta; Frutos, Roger; Zhou, Shuisen; Manguin, Sylvie

2017-11-01

China is approaching malaria elimination; however, well-documented information on malaria vectors is still missing, which could hinder the development of appropriate surveillance strategies and WHO certification. This review summarizes the nationwide distribution of malaria vectors, their bionomic characteristics, control measures, and related studies. After several years of effort, the area of distribution of the principal malaria vectors was reduced, in particular for Anopheles lesteri (synonym: An. anthropophagus) and Anopheles dirus s.l., which nearly disappeared from their former endemic regions. Anopheles sinensis is becoming the predominant species in southwestern China. The bionomic characteristics of these species have changed, and resistance to insecticides was reported. There is a need to update surveillance tools and investigate the role of secondary vectors in malaria transmission. Copyright © 2017 Elsevier Ltd. All rights reserved.

Differential susceptibilities of Anopheles albimanus and Anopheles pseudopunctipennis to infections with coindigenous Plasmodium vivax variants VK210 and VK247 in southern Mexico.

PubMed

Gonzalez-Ceron, L; Rodriguez, M H; Nettel, J C; Villarreal, C; Kain, K C; Hernandez, J E

1999-01-01

The susceptibilities to coindigenous Plasmodium vivax of colonized Anopheles albimanus and Anopheles pseudopunctipennis from southern Mexico were investigated by simultaneous feeding with infected blood obtained from patients. The genes encoding circumsporozoite protein variant types (VK210 and VK247) in blood samples were determined by PCR and oligonucleotide probe hybridization. A. albimanus was more susceptible to VK210, and A. pseudopunctipennis was more susceptible to VK247.

Standardizing operational vector sampling techniques for measuring malaria transmission intensity: evaluation of six mosquito collection methods in western Kenya.

PubMed

Wong, Jacklyn; Bayoh, Nabie; Olang, George; Killeen, Gerry F; Hamel, Mary J; Vulule, John M; Gimnig, John E

2013-04-30

Operational vector sampling methods lack standardization, making quantitative comparisons of malaria transmission across different settings difficult. Human landing catch (HLC) is considered the research gold standard for measuring human-mosquito contact, but is unsuitable for large-scale sampling. This study assessed mosquito catch rates of CDC light trap (CDC-LT), Ifakara tent trap (ITT), window exit trap (WET), pot resting trap (PRT), and box resting trap (BRT) relative to HLC in western Kenya to 1) identify appropriate methods for operational sampling in this region, and 2) contribute to a larger, overarching project comparing standardized evaluations of vector trapping methods across multiple countries. Mosquitoes were collected from June to July 2009 in four districts: Rarieda, Kisumu West, Nyando, and Rachuonyo. In each district, all trapping methods were rotated 10 times through three houses in a 3 × 3 Latin Square design. Anophelines were identified by morphology and females classified as fed or non-fed. Anopheles gambiae s.l. were further identified as Anopheles gambiae s.s. or Anopheles arabiensis by PCR. Relative catch rates were estimated by negative binomial regression. When data were pooled across all four districts, catch rates (relative to HLC indoor) for An. gambiae s.l (95.6% An. arabiensis, 4.4% An. gambiae s.s) were high for HLC outdoor (RR = 1.01), CDC-LT (RR = 1.18), and ITT (RR = 1.39); moderate for WET (RR = 0.52) and PRT outdoor (RR = 0.32); and low for all remaining types of resting traps (PRT indoor, BRT indoor, and BRT outdoor; RR < 0.08 for all). For Anopheles funestus, relative catch rates were high for ITT (RR = 1.21); moderate for HLC outdoor (RR = 0.47), CDC-LT (RR = 0.69), and WET (RR = 0.49); and low for all resting traps (RR < 0.02 for all). At finer geographic scales, however, efficacy of each trap type varied from district to district. ITT, CDC-LT, and WET appear to be effective methods for large-scale vector sampling in

Land use and land cover changes and spatiotemporal dynamics of anopheline larval habitats during a four-year period in a highland community of Africa.

PubMed

Munga, Stephen; Yakob, Laith; Mushinzimana, Emmanuel; Zhou, Guofa; Ouna, Tom; Minakawa, Noboru; Githeko, Andrew; Yan, Guiyun

2009-12-01

Spatial and temporal variations in the distribution of anopheline larval habitats and land use and land cover (LULC) changes can influence malaria transmission intensity. This information is important for understanding the environmental determinants of malaria transmission heterogeneity, and it is critical to the study of the effects of environmental changes on malaria transmission. In this study, we investigated the spatial and temporal variations in the distribution of anopheline larval habitats and LULC changes in western Kenya highlands over a 4-year period. Anopheles gambiae complex larvae were mainly confined to valley bottoms during both the dry and wet seasons. Although An. gambiae larvae were located in man-made habitats where riparian forests and natural swamps had been cleared, Anopheles funestus larvae were mainly found in permanent habitats in pastures. The association between land cover type and occurrence of anopheline larvae was statistically significant. The distribution of anopheline positive habitats varied significantly between months, during the survey. In 2004, the mean density of An. gambiae was significantly higher during the month of May, whereas the density of An. funestus peaked significantly in February. Over the study period, major LULC changes occurred mostly in the valley bottoms. Overall, farmland increased by 3.9%, whereas both pastures and natural swamps decreased by 8.9% and 20.9%, respectively. The area under forest cover was decreased by 5.8%. Land-use changes in the study area are favorable to An. gambiae larval development, thereby risking a more widespread distribution of malaria vector habitats and potentially increasing malaria transmission in western Kenya highlands.

Plasmodium falciparum Malaria, Southern Algeria, 2007

PubMed Central

Gassen, Ibrahim; Khechache, Yacine; Lamali, Karima; Tchicha, Boualem; Brengues, Cécile; Menegon, Michela; Severini, Carlo; Fontenille, Didier; Harrat, Zoubir

2010-01-01

An outbreak of Plasmodium falciparum malaria occurred in Tinzaouatine in southern Algeria in 2007. The likely vector, Anopheles gambiae mosquitoes, had not been detected in Algeria. Genes for resistance to chloroquine were detected in the parasite. The outbreak shows the potential for an increase in malaria vectors in Algeria. PMID:20113565

Differential Susceptibilities of Anopheles albimanus and Anopheles pseudopunctipennis to Infections with Coindigenous Plasmodium vivax Variants VK210 and VK247 in Southern Mexico

PubMed Central

Gonzalez-Ceron, Lilia; Rodriguez, Mario H.; Nettel, Jose C.; Villarreal, Cuauhtemoc; Kain, Kevin C.; Hernandez, Juan E.

1999-01-01

The susceptibilities to coindigenous Plasmodium vivax of colonized Anopheles albimanus and Anopheles pseudopunctipennis from southern Mexico were investigated by simultaneous feeding with infected blood obtained from patients. The genes encoding circumsporozoite protein variant types (VK210 and VK247) in blood samples were determined by PCR and oligonucleotide probe hybridization. A. albimanus was more susceptible to VK210, and A. pseudopunctipennis was more susceptible to VK247. PMID:9864243

Distinct population structure for co-occurring Anopheles goeldii and Anopheles triannulatus in Amazonian Brazil

PubMed Central

McKeon, Sascha Naomi; Moreno, Marta; Sallum, Maria Anise; Povoa, Marinete Marins; Conn, Jan Evelyn

2013-01-01

To evaluate whether environmental heterogeneity contributes to the genetic heterogeneity in Anopheles triannulatus, larval habitat characteristics across the Brazilian states of Roraima and Pará and genetic sequences were examined. A comparison with Anopheles goeldii was utilised to determine whether high genetic diversity was unique to An. triannulatus. Student t test and analysis of variance found no differences in habitat characteristics between the species. Analysis of population structure of An. triannulatus and An. goeldii revealed distinct demographic histories in a largely overlapping geographic range. Cytochrome oxidase I sequence parsimony networks found geographic clustering for both species; however nuclear marker networks depicted An. triannulatus with a more complex history of fragmentation, secondary contact and recent divergence. Evidence of Pleistocene expansions suggests both species are more likely to be genetically structured by geographic and ecological barriers than demography. We hypothesise that niche partitioning is a driving force for diversity, particularly in An. triannulatus. PMID:23903977

Social and cultural barriers to husbands' involvement in maternal health in rural Gambia.

PubMed

Lowe, Mat

2017-01-01

While many studies have documented a number of socio-cultural barriers to male involvement in maternal health, in The Gambia very little information is known about the social and cultural practices that characterized male involvement in maternal health. This study aims to explore some of the underlying social and cultural factors affecting husbands' involvement in maternal health issues pertaining to pregnancy and delivery in rural Gambia. Five focus group discussions and six in-depth interviews were conducted among rural men and traditional birth attendants in five areas of rural Gambia. The discussion was directed to the roles of male partners in pregnancy and delivery and the difficulties they face regarding taking care of their wives. The data resulting from the discussion was audio-recorded, transcribed verbatim, and analyzed thematically. In general, rural Gambian men and traditional birth attendants (TBAs) reported that husbands' involvement in maternal health is highly desirable, but is influenced by many factors, such as the traditional conceptualization associated with pregnancy and delivery as women's domain. In addition, many men do not believe that pregnancy chores warrant their efforts compared to other competing social responsibilities. This issue may be more complicated in polygamous marriages where there is rivalry among co-wives and in neighborhoods where men who help with house chores may be subjected to mockery. These findings suggest that husbands' involvement in maternal health in The Gambia is influenced by the prevailing social and cultural practices of gender role and norms, which are also at the root of maternal health problems.

Infertility in the Gambia: Traditional and Modern Health Care.

ERIC Educational Resources Information Center

Sundby, Johanne

1997-01-01

A population survey was undertaken to study infertility in Gambia. All infertile women in 24 randomly selected enumeration areas were assessed. Problems faced, coping mechanisms employed, and types of health care available were examined. Patterns of consultation with traditional versus formal health care and rural/urban differences were uncovered…

Laboratory Evaluations of the Fractions Efficacy of Annona senegalensis (Annonaceae) Leaf Extract on Immature Stage Development of Malarial and Filarial Mosquito Vectors

PubMed Central

Lame, Younoussa; Nukenine, Elias Nchiwan; Pierre, Danga Yinyang Simon; Elijah, Ajaegbu Eze; Esimone, Charles Okechukwu

2015-01-01

Background: Within the framework to control mosquitoes, ovicidal, larvicidal and pupicidal activity of Annona senegalensis leaf extract and its 4 fractions against Anopheles gambiae and Culex quinquefasciatus were evaluated in the laboratory conditions. Methods: Ovicidal test was performed by submitting at least 100 eggs of mosquitoes to 125, 250, 500, 1000 and 2000 ppm concentrations, while larvicidal and pupicidal effects were assessed by submitting 25 larvae or pupae to the concentrations of 2500, 1250, 625 and 312.5 ppm of plant extract or fractions of A. senegalensis. Results: The eggs of An. gambiae were most affected by N-hexane (0.00% hatchability) and chloroform (03.67% hatchability) fractions compared to Cx. quinquefasciatus where at least 25 % hatchability were recorded at 2000 ppm. For larvicidal test, N-hexane (LC50= 298.8 ppm) and chloroform (LC50= 418.3 ppm) fractions were more effective than other fractions on An. gambiae larvae while, a moderate effectiveness was also observed with N-hexane (LC50= 2087.6 ppm), chloroform (LC50= 9010.1 ppm) fractions on Cx. quinquefasciatus larvae. The highest mortality percent of the pupae were also recorded with N-hexane and chloroform fractions on An. gambiae at 2500 ppm. As for Cx. quinquefasciatus only 50 % and 36 % mortality were recorded with N-hexane and chloroform fractions respectively. Conclusion: The extract of A. senegalensis was toxic on immature stage of mosquito species tested. By splitting methanolic crude extract, only N-hexane and chloroform fractions were revealed to possess a mosquitocidal effects and could be considered and utilized for future immature mosquito vectors control. PMID:26623434

Larval habitats of Anopheles species in a rural settlement on the malaria frontier of southwest Amazon, Brazil.

PubMed

Rufalco-Moutinho, Paulo; Schweigmann, Nicolás; Bergamaschi, Denise Pimentel; Mureb Sallum, Maria Anice

2016-12-01

Rural settlements are social arrangements expanding in the Amazon region, which generate disturbances in the natural environment, thus affecting the ecology of the species of Anopheles and thus the malaria transmission. Larval habitats are important sources for maintenance of mosquito vector populations, and holding back a natural watercourse is a usual process in the establishment of rural settlements, since the formation of micro-dams represents a water resource for the new settlers. Identifying characteristics of the larval habitats that may be associated with both the presence and abundance of Anopheles vectors species in an environment under ecological transition is background for planning vector control strategies in rural areas in the Amazon. Anopheles larvae collections were performed in two major types of habitats: natural and flow-limited water collections that were constructed by holding back the original watercourse. A total of 3123 Anopheles spp. larvae were captured in three field-sampling collections. The majority of the larvae identified were taken from flow-limited water collections belonged to species of the Nyssorhynchus subgenus (92%), whereas in the natural larval habitats a fewer number of individuals belonged to the Stethomyia (5%) and Anopheles (3%) subgenera. The total of Nyssorhynchus identified (1818), 501 specimens belonged to An. darlingi, 750 to An. triannulatus and 567 for others remaining species. In addition, 1152 could not be identified to subgenus/species level, because they were either in the first-instar or damaged. The primary vector in areas of the Amazon river basin, An. darlingi, was found exclusively in man-made habitats. Statistical analysis display An. triannulatus with specialist behavior for characteristics of man-made habitats. Modifications in the natural environment facilitate the rise of larval habitats for species with epidemiological importance for malaria in the region. This study showed that man-made habitats

Ion and solute transport by prestin in Drosophila and Anopheles

PubMed Central

Hirata, Taku; Czapar, Anna; Brin, Lauren R.; Haritonova, Alyona; Bondeson, Daniel P.; Linser, Paul J.; Cabrero, Pablo; Dow, Julian A. T.; Romero, Michael F.

2012-01-01

The gut and Malpighian tubules of insects are the primary sites of active solute and water transport for controlling hemolymph and urine composition, pH, and osmolarity. These processes depend on ATPase (pumps), channels and solute carriers (Slc proteins). Maturation of genomic databases enables us to identify the putative molecular players for these processes. Anion transporters of the Slc4 family, AE1 and NDAE1, have been reported as HCO3− transporters, but are only part of the story. Here we report Dipteran (Drosophila melanogaster (d) and Anopheles gambiae (Ag)) anion exchangers, belonging to the Slc26 family, which are multi-functional anion exchangers. One Drosophila and two Ag homologues of mammalian Slc26a5 (prestin) and Slc26a6 (aka, PAT1, CFEX) were identified and designated dPrestin, AgPrestinA and AgPrestinB. dPrestin and AgPrestinB show electrogenic anion exchange (Cl−/nHCO3−, Cl−/SO42− and Cl−/oxalate2−) in an oocyte expression system. Since these transporters are the only Dipteran Slc26 proteins whose transport is similar to mammalian Slc26a6, we submit that Dipteran Prestin are functional and even molecular orthologues of mammalian Slc26a6. OSR1 kinase increases dPrestin ion transport, implying another set of physiological processes controlled by WNK/SPAK signaling in epithelia. All of these mRNAs are highly expressed in the gut and Malpighian tubules. Dipteran Prestin proteins appear suited for central roles in bicarbonate, sulfate and oxalate metabolism including generating the high pH conditions measured in the Dipteran midgut lumen. Finally, we present and discuss Drosophila genetic models that integrate these processes. PMID:22321763

Economic evaluation of pneumococcal conjugate vaccination in The Gambia.

PubMed

Kim, Sun-Young; Lee, Gene; Goldie, Sue J

2010-09-03

Gambia is the second GAVI support-eligible country to introduce the 7-valent pneumococcal conjugate vaccine (PCV7), but a country-specific cost-effectiveness analysis of the vaccine is not available. Our objective was to assess the potential impact of PCVs of different valences in The Gambia. We synthesized the best available epidemiological and cost data using a state-transition model to simulate the natural histories of various pneumococcal diseases. For the base-case, we estimated incremental cost (in 2005 US dollars) per disability-adjusted life year (DALY) averted under routine vaccination using PCV9 compared to no vaccination. We extended the base-case results for PCV9 to estimate the cost-effectiveness of PCV7, PCV10, and PCV13, each compared to no vaccination. To explore parameter uncertainty, we performed both deterministic and probabilistic sensitivity analyses. We also explored the impact of vaccine efficacy waning, herd immunity, and serotype replacement, as a part of the uncertainty analyses, by assuming alternative scenarios and extrapolating empirical results from different settings. Assuming 90% coverage, a program using a 9-valent PCV (PCV9) would prevent approximately 630 hospitalizations, 40 deaths, and 1000 DALYs, over the first 5 years of life of a birth cohort. Under base-case assumptions ($3.5 per vaccine), compared to no intervention, a PCV9 vaccination program would cost $670 per DALY averted in The Gambia. The corresponding values for PCV7, PCV10, and PCV13 were $910, $670, and $570 per DALY averted, respectively. Sensitivity analyses that explored the implications of the uncertain key parameters showed that model outcomes were most sensitive to vaccine price per dose, discount rate, case-fatality rate of primary endpoint pneumonia, and vaccine efficacy against primary endpoint pneumonia. Based on the information available now, infant PCV vaccination would be expected to reduce pneumococcal diseases caused by S. pneumoniae in The Gambia

Urban agriculture and Anopheles habitats in Dar es Salaam, Tanzania.

PubMed

Dongus, Stefan; Nyika, Dickson; Kannady, Khadija; Mtasiwa, Deo; Mshinda, Hassan; Gosoniu, Laura; Drescher, Axel W; Fillinger, Ulrike; Tanner, Marcel; Killeen, Gerry F; Castro, Marcia C

2009-05-01

A cross-sectional survey of agricultural areas, combined with routinely monitored mosquito larval information, was conducted in urban Dar es Salaam, Tanzania, to investigate how agricultural and geographical features may influence the presence of Anopheles larvae. Data were integrated into a geographical information systems framework, and predictors of the presence of Anopheles larvae in farming areas were assessed using multivariate logistic regression with independent random effects. It was found that more than 5% of the study area (total size 16.8 km2) was used for farming in backyard gardens and larger open spaces. The proportion of habitats containing Anopheles larvae was 1.7 times higher in agricultural areas compared to other areas (95% confidence interval = 1.56-1.92). Significant geographic predictors of the presence of Anopheles larvae in gardens included location in lowland areas, proximity to river, and relatively impermeable soils. Agriculture-related predictors comprised specific seedbed types, mid-sized gardens, irrigation by wells, as well as cultivation of sugar cane or leafy vegetables. Negative predictors included small garden size, irrigation by tap water, rainfed production and cultivation of leguminous crops or fruit trees. Although there was an increased chance of finding Anopheles larvae in agricultural sites, it was found that breeding sites originated by urban agriculture account for less than a fifth of all breeding sites of malaria vectors in Dar es Salaam. It is suggested that strategies comprising an integrated malaria control effort in malaria-endemic African cities include participatory involvement of farmers by planting shade trees near larval habitats.

Climate change and altitudinal structuring of malaria vectors in south-western Cameroon: their relation to malaria transmission.

PubMed

Tanga, M C; Ngundu, W I; Judith, N; Mbuh, J; Tendongfor, N; Simard, Frédéric; Wanji, S

2010-07-01

An entomological survey was conducted in Cameroon between October 2004 and September 2005, in nine localities targeted for malaria vector control based on adult productivity and variability. Mosquitoes were collected by human-landing catches (HLCs) and pyrethrum spray catches. A total of 12 500 anophelines were collected and dissected: Anopheles gambiae s.l. (56.86%), An. funestus s.l. (32.57%), An. hancocki (9.38%), and An. nili (1.18%). Applying PCR revealed that specimens of the An. funestus group were An. funestus s.s. and An. gambiae complex were mostly An. melas and An. gambiae s.s. of the M and S molecular forms with the M forms being the most predominant. The natural distribution patterns of Anopheles species were largely determined by altitude with some species having unique environmental tolerance limits. A human blood index (HBI) of 99.05% was recorded. Mean probability of daily survival of the malaria vectors was 0.92, with annual mean life expectancy of 21.9 days and the expectation of infective life was long with a mean of 7.4 days. The high survival rates suggest a high vector potential for the species. This information enhances the development of a more focused and informed vector control intervention. Copyright 2010 Royal Society of Tropical Medicine and Hygiene. Published by Elsevier Ltd. All rights reserved.

Innexin AGAP001476 Is Critical for Mediating Anti-Plasmodium Responses in Anopheles Mosquitoes

PubMed Central

Li, Michelle W. M.; Wang, Jiuling; Zhao, Yang O.; Fikrig, Erol

2014-01-01

The Toll and IMD pathways are known to be induced upon Plasmodium berghei and Plasmodium falciparum infection, respectively. It is unclear how Plasmodium or other pathogens in the blood meal and their invasion of the midgut epithelium would trigger the innate immune responses in immune cells, in particular hemocytes. Gap junctions, which can mediate both cell-to-cell and cell-to-extracellular communication, may participate in this signal transduction. This study examined whether innexins, gap junction proteins in insects, are involved in anti-Plasmodium responses in Anopheles gambiae. Inhibitor studies using carbenoxolone indicated that blocking innexons resulted in an increase in Plasmodium oocyst number and infection prevalence. This was accompanied by a decline in TEP1 levels in carbenoxolone-treated mosquitoes. Innexin AGAP001476 mRNA levels in midguts were induced during Plasmodium infection and a knockdown of AGAP001476, but not AGAP006241, caused an induction in oocyst number. Silencing AGAP001476 caused a concurrent increase in vitellogenin levels, a TEP1 inhibitor, in addition to a reduced level of TEP1-LRIM1-APL1C complex in hemolymph. Both vitellogenin and TEP1 are regulated by Cactus under the Toll pathway. Simultaneous knockdown of cactus and AGAP001476 failed to reverse the near refractoriness induced by the knockdown of cactus, suggesting that the AGAP001476-mediated anti-Plasmodium response is Cactus-dependent. These data demonstrate a critical role for innexin AGAP001476 in mediating innate immune responses against Plasmodium through Toll pathway in mosquitoes. PMID:25035430

GNBP domain of Anopheles darlingi: are polymorphic inversions and gene variation related to adaptive evolution?

PubMed

Bridi, L C; Rafael, M S

2016-02-01

Anopheles darlingi is the main malaria vector in humans in South America. In the Amazon basin, it lives along the banks of rivers and lakes, which responds to the annual hydrological cycle (dry season and rainy season). In these breeding sites, the larvae of this mosquito feed on decomposing organic and microorganisms, which can be pathogenic and trigger the activation of innate immune system pathways, such as proteins Gram-negative binding protein (GNBP). Such environmental changes affect the occurrence of polymorphic inversions especially at the heterozygote frequency, which confer adaptative advantage compared to homozygous inversions. We mapped the GNBP probe to the An. darlingi 2Rd inversion by fluorescent in situ hybridization (FISH), which was a good indicator of the GNBP immune response related to the chromosomal polymorphic inversions and adaptative evolution. To better understand the evolutionary relations and time of divergence of the GNBP of An. darlingi, we compared it with nine other mosquito GNBPs. The results of the phylogenetic analysis of the GNBP sequence between the species of mosquitoes demonstrated three clades. Clade I and II included the GNBPB5 sequence, and clade III the sequence of GNBPB1. Most of these sequences of GNBP analyzed were homologous with that of subfamily B, including that of An. gambiae (87 %), therefore suggesting that GNBP of An. darling belongs to subfamily B. This work helps us understand the role of inversion polymorphism in evolution of An. darlingi.

Paratransgenesis to control malaria vectors: a semi-field pilot study.

PubMed

Mancini, Maria Vittoria; Spaccapelo, Roberta; Damiani, Claudia; Accoti, Anastasia; Tallarita, Mario; Petraglia, Elisabetta; Rossi, Paolo; Cappelli, Alessia; Capone, Aida; Peruzzi, Giulia; Valzano, Matteo; Picciolini, Matteo; Diabaté, Abdoulaye; Facchinelli, Luca; Ricci, Irene; Favia, Guido

2016-03-10

Malaria still remains a serious health burden in developing countries, causing more than 1 million deaths annually. Given the lack of an effective vaccine against its major etiological agent, Plasmodium falciparum, and the growing resistance of this parasite to the currently available drugs repertoire and of Anopheles mosquitoes to insecticides, the development of innovative control measures is an imperative to reduce malaria transmission. Paratransgenesis, the modification of symbiotic organisms to deliver anti-pathogen effector molecules, represents a novel strategy against Plasmodium development in mosquito vectors, showing the potential to reduce parasite development. However, the field application of laboratory-based evidence of paratransgenesis imposes the use of more realistic confined semi-field environments. Large cages were used to evaluate the ability of bacteria of the genus Asaia expressing green fluorescent protein (Asaia (gfp)), to diffuse in Anopheles stephensi and Anopheles gambiae target mosquito populations. Asaia (gfp) was introduced in large cages through the release of paratransgenic males or by sugar feeding stations. Recombinant bacteria transmission was directly detected by fluorescent microscopy, and further assessed by molecular analysis. Here we show the first known trial in semi-field condition on paratransgenic anophelines. Modified bacteria were able to spread at high rate in different populations of An. stephensi and An. gambiae, dominant malaria vectors, exploring horizontal ways and successfully colonising mosquito midguts. Moreover, in An. gambiae, vertical and trans-stadial diffusion mechanisms were demonstrated. Our results demonstrate the considerable ability of modified Asaia to colonise different populations of malaria vectors, including pecies where its association is not primary, in large environments. The data support the potential to employ transgenic Asaia as a tool for malaria control, disclosing promising perspective

Comparison of the standard WHO susceptibility tests and the CDC bottle bioassay for the determination of insecticide susceptibility in malaria vectors and their correlation with biochemical and molecular biology assays in Benin, West Africa.

PubMed

Aïzoun, Nazaire; Ossè, Razaki; Azondekon, Roseric; Alia, Roland; Oussou, Olivier; Gnanguenon, Virgile; Aikpon, Rock; Padonou, Gil Germain; Akogbéto, Martin

2013-05-20

The detection of insecticide resistance in natural populations of Anopheles vectors is absolutely necessary for malaria control. In the African region, the WHO insecticide susceptibility test is the most common method for assessing resistance status. In order to search for a simple, rapid and more reliable technique in the assessment of insecticide resistance in malaria vectors, we compared the WHO tests with the CDC bottle bioassay in the Ouemé province of southern Benin where insecticide resistance has been widely reported. Larvae and pupae of Anopheles gambiae s.l. mosquitoes were collected from the breeding sites in Ouemé. WHO and CDC susceptibility tests were conducted simultaneously on unfed female mosquitoes aged 2-5 days old. WHO bioassays were performed with impregnated papers of deltamethrin (0.05%) and bendiocarb (0.1%), whereas CDC bioassays were performed with stock solutions of deltamethrin (12.5 μg per bottle) and bendiocarb (12.5 μg per bottle). PCR techniques were used to detect species, Kdr and Ace-1 mutations. CDC biochemical assays using synergists were also conducted to assess the metabolic resistance. A slight decrease in mortality rates was observed with 97.95% and 98.33% obtained from CDC and WHO bioassays respectively in populations of mosquitoes from Adjara and Dangbo. PCR revealed that all specimens tested were Anopheles gambiae s.s. The Kdr mutation was found at high frequency in all populations and both the Kdr mutation and mono-oxygenase enzymes were implicated as mechanisms of pyrethroid resistance in An. gambiae from Misserete. This study emphasizes that both WHO and CDC bioassays give similar results with regards to the susceptibility of mosquitoes to insecticides in southern Benin. There were complementarities between both methods, however, some specificity was noted for each of the two methods used. Both Kdr and metabolic mechanisms were implicated in the resistance.

Good performances but short lasting efficacy of Actellic 50 EC Indoor Residual Spraying (IRS) on malaria transmission in Benin, West Africa

PubMed Central

2014-01-01

Background The National Malaria Control Program (NMCP) has been using pirimiphos methyl for the first time for indoor residual spraying (IRS) in Benin. The first round was a success with a significant decrease of entomological indicators of malaria transmission in the treated districts. We present the results of the entomological impact on malaria transmission. Entomologic parameters in the control area were compared with those in intervention sites. Methods Mosquito collections were carried out in three districts in the Atacora-Dongo region of which two were treated with pirimiphos methyl (Actellic 50EC) (Tanguiéta and Kouandé) and the untreated (Copargo) served as control. Anopheles gambiae s.l. populations were sampled monthly by human landing catch. In addition, window exit traps and pyrethrum spray catches were performed to assess exophagic behavior of Anopheles vectors. In the three districts, mosquito collections were organized to follow the impact of pirimiphos methyl IRS on malaria transmission and possible changes in the behavior of mosquitoes. The residual activity of pirimiphos methyl in the treated walls was also assessed using WHO bioassay test. Results A significant reduction (94.25%) in human biting rate was recorded in treated districts where an inhabitant received less than 1 bite of An. gambiae per night. During this same time, the entomological inoculation rate (EIR) dramatically declined in the treated area (99.24% reduction). We also noted a significant reduction in longevity of the vectors and an increase in exophily induced by pirimiphos methyl on An. gambiae. However, no significant impact was found on the blood feeding rate. Otherwise, the low residual activity of Actellic 50 EC, which is three months, is a disadvantage. Conclusion Pirimiphos methyl was found to be effective for IRS in Benin. However, because of the low persistence of Actellic 50EC used in this study on the treated walls, the recourse to another more residual formulation

Factors influencing the spatial distribution of Anopheles larvae in Coimbatore District, Tamil Nadu, India.

PubMed

Arjunan, Naresh Kumar; Kadarkarai, Murugan; Kumar, Shobana; Pari, Madhiyazhagan; Thiyagarajan, Nataraj; Vincent, C Thomas; Barnard, Donald R

2015-12-01

Malaria causes extensive morbidity and mortality in humans and results in significant economic losses in India. The distribution of immature malaria-transmitting Anopheles mosquitoes was studied in 17 villages in Coimbatore District as a prelude to the development and implementation of vector control strategies that are intended to reduce the risk of human exposure to potentially infectious mosquitoes. Eight Anopheles species were recorded. The most numerous species were Anopheles vagus, Anopheles subpictus, and Anopheles hyrcanus. The location of mosquito development sites and the density of larvae in each village was evaluated for correlation with selected demographic, biologic, and land use parameters using remote sensing and geographic information systems (GIS) technology. We found the number of mosquito development sites in a village and the density of larvae in such sites to be positively correlated with human population density but not the surface area (km(2)) of the village. The number of mosquito development sites and the density of larvae in each site were not correlated. Data from this study are being used to construct a GIS-based mapping system that will enable the location of aquatic habitats with Anopheles larvae in the Coimbatore District, Tamil Nadu, India as target sites for the application of vector control. Copyright © 2015 Elsevier B.V. All rights reserved.

A Bioinformatics Approach for Integrated Transcriptomic and Proteomic Comparative Analyses of Model and Non-sequenced Anopheline Vectors of Human Malaria Parasites*

PubMed Central

Mohien, Ceereena Ubaida; Colquhoun, David R.; Mathias, Derrick K.; Gibbons, John G.; Armistead, Jennifer S.; Rodriguez, Maria C.; Rodriguez, Mario Henry; Edwards, Nathan J.; Hartler, Jürgen; Thallinger, Gerhard G.; Graham, David R.; Martinez-Barnetche, Jesus; Rokas, Antonis; Dinglasan, Rhoel R.

2013-01-01

Malaria morbidity and mortality caused by both Plasmodium falciparum and Plasmodium vivax extend well beyond the African continent, and although P. vivax causes between 80 and 300 million severe cases each year, vivax transmission remains poorly understood. Plasmodium parasites are transmitted by Anopheles mosquitoes, and the critical site of interaction between parasite and host is at the mosquito's luminal midgut brush border. Although the genome of the “model” African P. falciparum vector, Anopheles gambiae, has been sequenced, evolutionary divergence limits its utility as a reference across anophelines, especially non-sequenced P. vivax vectors such as Anopheles albimanus. Clearly, technologies and platforms that bridge this substantial scientific gap are required in order to provide public health scientists with key transcriptomic and proteomic information that could spur the development of novel interventions to combat this disease. To our knowledge, no approaches have been published that address this issue. To bolster our understanding of P. vivax–An. albimanus midgut interactions, we developed an integrated bioinformatic-hybrid RNA-Seq-LC-MS/MS approach involving An. albimanus transcriptome (15,764 contigs) and luminal midgut subproteome (9,445 proteins) assembly, which, when used with our custom Diptera protein database (685,078 sequences), facilitated a comparative proteomic analysis of the midgut brush borders of two important malaria vectors, An. gambiae and An. albimanus. PMID:23082028

A bioinformatics approach for integrated transcriptomic and proteomic comparative analyses of model and non-sequenced anopheline vectors of human malaria parasites.

PubMed

Ubaida Mohien, Ceereena; Colquhoun, David R; Mathias, Derrick K; Gibbons, John G; Armistead, Jennifer S; Rodriguez, Maria C; Rodriguez, Mario Henry; Edwards, Nathan J; Hartler, Jürgen; Thallinger, Gerhard G; Graham, David R; Martinez-Barnetche, Jesus; Rokas, Antonis; Dinglasan, Rhoel R

2013-01-01

Malaria morbidity and mortality caused by both Plasmodium falciparum and Plasmodium vivax extend well beyond the African continent, and although P. vivax causes between 80 and 300 million severe cases each year, vivax transmission remains poorly understood. Plasmodium parasites are transmitted by Anopheles mosquitoes, and the critical site of interaction between parasite and host is at the mosquito's luminal midgut brush border. Although the genome of the "model" African P. falciparum vector, Anopheles gambiae, has been sequenced, evolutionary divergence limits its utility as a reference across anophelines, especially non-sequenced P. vivax vectors such as Anopheles albimanus. Clearly, technologies and platforms that bridge this substantial scientific gap are required in order to provide public health scientists with key transcriptomic and proteomic information that could spur the development of novel interventions to combat this disease. To our knowledge, no approaches have been published that address this issue. To bolster our understanding of P. vivax-An. albimanus midgut interactions, we developed an integrated bioinformatic-hybrid RNA-Seq-LC-MS/MS approach involving An. albimanus transcriptome (15,764 contigs) and luminal midgut subproteome (9,445 proteins) assembly, which, when used with our custom Diptera protein database (685,078 sequences), facilitated a comparative proteomic analysis of the midgut brush borders of two important malaria vectors, An. gambiae and An. albimanus.

Effect of two different house screening interventions on exposure to malaria vectors and on anaemia in children in The Gambia: a randomised controlled trial.

PubMed

Kirby, Matthew J; Ameh, David; Bottomley, Christian; Green, Clare; Jawara, Musa; Milligan, Paul J; Snell, Paul C; Conway, David J; Lindsay, Steve W

2009-09-19

House screening should protect people against malaria. We assessed whether two types of house screening--full screening of windows, doors, and closing eaves, or installation of screened ceilings--could reduce house entry of malaria vectors and frequency of anaemia in children in an area of seasonal malaria transmission. During 2006 and 2007, 500 occupied houses in and near Farafenni town in The Gambia, an area with low use of insecticide-treated bednets, were randomly assigned to receive full screening, screened ceilings, or no screening (control). Randomisation was done by computer-generated list, in permuted blocks of five houses in the ratio 2:2:1. Screening was not treated with insecticide. Exposure to mosquitoes indoors was assessed by fortnightly light trap collections during the transmission season. Primary endpoints included the number of female Anopheles gambiae sensu lato mosquitoes collected per trap per night. Secondary endpoints included frequency of anaemia (haemoglobin concentration <80 g/L) and parasitaemia at the end of the transmission season in children (aged 6 months to 10 years) who were living in the study houses. Analysis was by modified intention to treat (ITT), including all randomised houses for which there were some outcome data and all children from those houses who were sampled for haemoglobin and parasitaemia. This study is registered as an International Standard Randomised Controlled Trial, number ISRCTN51184253. 462 houses were included in the modified ITT analysis (full screening, n=188; screened ceilings, n=178; control, n=96). The mean number of A gambiae caught in houses without screening was 37.5 per trap per night (95% CI 31.6-43.3), compared with 15.2 (12.9-17.4) in houses with full screening (ratio of means 0.41, 95% CI 0.31-0.54; p<0.0001) and 19.1 (16.1-22.1) in houses with screened ceilings (ratio 0.53, 0.40-0.70; p<0.0001). 755 children completed the study, of whom 731 had complete clinical and covariate data and were

Hybrid sterility in crosses between two Brazilian sibling species of the Anopheles albitarsis complex.

PubMed

Fontoura, Nathalia Giglio; Araki, Alejandra Saori; Van Der Maas Azevedo, Renata; Galardo, Allan Kardec Ribeiro; Peixoto, Alexandre Afranio; Lima, José Bento Pereira

2014-12-04

Complexes of cryptic species are common in several taxa and this is also the case in the Anopheles genus, a group including all known human malaria vectors. The Anopheles albitarsis complex comprises at least nine cryptic species, some of which are implicated as vectors of human malaria. Several different types of data have been generated for this species complex such as cytogenetics, alloenzymes, morphological and feeding behavioral, hybridization experiments, RAPD-PCR and RFLP and mitochondrial and nuclear markers. Studies focused on its postzygotic isolation are still somewhat rare in the literature despite their importance to understand the speciation process and the level of gene flow potentially occurring among the different sibling species. Hybridization experiments between Anopheles albitarsis s.s. and Anopheles marajoara, as well as backcrosses between hybrids and Anopheles albitarsis s.s., were performed using the induced mating technique. Results were compared to intraspecific crosses. Larva-to-adult viability and sex ratio were also assessed. Male hybrids show very low insemination rates and nearly complete sterility, apparently due to abnormalities in their reproductive organs. Evidence of partial sterility among the hybrid females was also observed. Our data indicated that Anopheles albitarsis s.s. and Anopheles marajoara show a high level of postzygotic isolation with a strong hybrid male sterility. This result is consistent with the Haldane's rule which states that in interspecific crosses the heterogametic sex is the first to be affected. However, the fact that the females are not completely sterile raises the possibility of introgression between these two siblings species.

Mark-recapture studies of host selection by Anopheles (Anopheles) vestitipennis.

PubMed

Ulloa, Armando; Arredondo-Jiménez, Juan I; Rodriguez, Mario H; Fernández-Salas, Ildefonso

2002-03-01

We present herein the results of a series of mark-recapture experiments with female Anopheles vestitipennis. Theses experiments used human and animal hosts to assess the degree of anthropophily of field-caught specimens, originally collected on either host, and of their offspring. Fidelity of mosquitoes to particular hosts was estimated by recapturing marked host-seeking mosquitoes returning for a 2nd blood meal. Results indicated that mosquitoes seeking animal hosts were more faithful (80.48%; 33 of 41) in returning to their original host than were those seeking human hosts (63%; 49 of 78).

[Preliminary investigation of four anopheles larvae samples susceptibility to chlorpyrifos in Tunisia].

PubMed

Krida, G; Bouattour, A; Rhaim, A; el Kebir, A; Jlidi, R

1998-01-01

Larvae susceptibility to chlorpyrifos is studied on four Tunisian Anopheles larvas samples: Anopheles labranchiae from Rades (South of Tunis) and Menchar (region of Beja), in the North of Tunisia, A. sergentii from Meknassy and A. multicolor from Sidi Bouzid, both in the Centre of Tunisia. The test results of larvae susceplibility indicate that the LC50 and the LC95 values are less than 0.002 and 0.02 mg l-1 respectively and their 95% confidence limits overlap. We also notice that the studied samples show the same susceptibility to chlorpyrifos. The results can be used as a base data further studies on the susceptibility of Anopheles to chemical insecticides.

Unexpected anthropophily in the potential secondary malaria vectors Anopheles coustani s.l. and Anopheles squamosus in Macha, Zambia.

PubMed

Fornadel, Christen M; Norris, Laura C; Franco, Veronica; Norris, Douglas E

2011-08-01

Anopheles coustani s.l. and Anopheles squamosus are sub-Saharan mosquito species that have been implicated in malaria transmission. Although generally believed to be of negligible importance due to their overwhelmingly zoophilic behavior, An. coustani s.l. and An. squamosus made up a large proportion of the anophelines collected by human landing catches during the 2007-2008 and 2008-2009 rainy seasons in Macha, Zambia. Further, polymerase chain reaction-based blood meal identification showed that the majority of blood meals from these mosquito species caught in human-baited Centers for Disease Control light traps were from human hosts. Although no An. coustani s.l. or An. squamosus were found to be positive for Plasmodium, the demonstrated anthropophilic tendencies of these mosquitoes in southern Zambia suggest their potential as secondary malaria vectors.

Morphological Analysis of Three Populations of Anopheles (Nyssorhynchus) Nuneztovari Gabaldon (Diptera: Culicidae) from Colombia

DTIC Science & Technology

2008-02-01

81. Gabaldon A 1940. Estudios sobre Anophelinos. Serie I. 1. Descrip- ción de Anopheles (Nyssorhynchus) nuneztovari n. sp. y consid- eraciones sobre...N 2007. Introducción al estudio Taxonómico de Anopheles de Colombia Claves y notas de distribución, 1st ed., Universidad del Valle, Cali, 237 pp...divergence in of Anopheles nuneztovari (Diptera: Culicidae) from Brazil and Colombia. Am J Trop Med Hyg 60: 1010-1018. Sutil OE 1976. Redescripción de la

New records of Anopheles (Diptera: Culicidae) in Puerto Carreño, Vichada, Colombia

PubMed Central

Brochero, Helena; Conn, Jan E.

2017-01-01

Malaria is endemic in Vichada department in the Orinoquia region of Colombia. Reports of species in the genus Anopheles Meigen 1818, responsible for transmission of Plasmodium spp. parasites leading to malaria cases, should result in strengthened entomological surveillance and a clearer focus on the most effective vector control strategies. Here we present new records of Anopheles (Anopheles) shannoni Davis, 1931 and A. (A.) costai & forattini for Vichada department, Colombia. Additionally, we have new records of A. (Nyssorhynchus) triannulatus Neiva & Pinto, 1922, and A. (A.) mattogrossensis Lutz & Neiva, 1911 for Puerto Carreño, the capital of Vichada department. PMID:29104672